Novel Compositions for Preventing and/or Treating Degenerative Disorders of the Central Nervous System

ABSTRACT

The present invention provides novel compounds as well as compositions and methods using the same for preventing and/or treating degenerative disorders of the central nervous system. In particular, the present invention provides methods for preventing and/or treating Parkinson&#39;s disease.

FIELD OF THE INVENTION

The present invention provides novel compounds, known as pharmacologicalchaperones, as well as compositions and methods using the same forpreventing and/or treating degenerative disorders of the central nervoussystem. In particular, the present invention provides methods forpreventing and/or treating Parkinson's disease.

BACKGROUND OF THE INVENTION

Many degenerative disorders of the central nervous system are associatedwith pathologic aggregation of proteins or lipids. For example,synucleinopathies are a group of diseases that arise from disruption ofsynuclein protein homeostasis. In particular, alpha-synucleinaggregation is associated with pathological conditions characterized byLewy bodies, such as Parkinson's disease, dementia with Lewy bodies, andmultiple system atrophy. Likewise, alpha-synuclein fragment, non-Abetacomponent, is found in amyloid plaques of Alzheimer's disease. Recently,enhancement of glucocerebrosidase (beta-glucosidase; GCase) activity inthe brain has been shown to prevent accumulation of synuclein in thebrain (Sean Clark, Ying Sun, You-Hai Xu, Gregory Grabowski, and BrandonWustman, “A biochemical link between Gaucher and Parkinson's disease anda potential new approach to treating synucleinopathies: apharmacological chaperone for beta-glucocerebrosidase preventsaccumulation of alpha-synuclein in a Parkinson's mouse model,” Presentedat the Society for Neuroscience Annual Meeting, San Diego, Calif.,2007). Thus, agents that enhance GCase activity may provide relief forpatients at risk for developing or diagnosed with degenerative disordersof the central nervous system.

There is a need for new therapeutic compounds that can be used toprevent and/or treat degenerative disorders of the central nervoussystem that provide patients with a higher quality of life and achieve abetter clinical outcome. In particular, there is a need for newtherapeutic compounds to prevent and/or treat synucleinopathies such asParkinson's disease and Alzheimer's disease that provide patients with ahigher quality of life and achieve a better clinical outcome.

SUMMARY OF THE INVENTION

The present invention provides novel compounds as well as compositionsand methods using the same to prevent and/or treat a degenerativedisorder of the central nervous system in a patient at risk fordeveloping or diagnosed with the same which includes administering tothe patient in need thereof an effective amount of a compound describedherein.

In one aspect, there is provided a compound as well as compositions andmethods using the same to prevent and/or treat a degenerative disorderof the central nervous system in a patient at risk for developing ordiagnosed with the same which includes administering to the patient inneed thereof an effective amount of a compound defined by Formula I:

wherein:

-   -   R¹ is C(R²)(R³)(R⁴);    -   R² is hydrogen, —OH or halogen;    -   R³ is hydrogen, —OH, halogen or C₁₋₈ alkyl;    -   R⁴ is halogen, C₁₋₈ alkyl, substituted C₁₋₈ alkyl, aryl,        substituted aryl, alkylcycloalkyl or substituted        alkylcycloalkyl;    -   R³ and R⁴ may join with the carbon to which they are attached to        form a cycloalkyl ring, which may be optionally substituted,        preferably with halogen and more preferably with one or more        fluorine atoms;    -   R⁶ is hydrogen, C₁₋₈ alkyl, substituted C₁₋₈ alkyl, arylalkyl,        substituted arylalkyl, alkylaryl, or substituted alkylaryl;    -   Z is optional, when present Z is —(CH₂)₁₋₈—, —C(═O) —,        —S(═O)₂NH—, —S(═O)₂—,    -   —C(═S)NH—, —S(═O)₂—CH₃, C(═O)—NH—, —S(═O)₂—NR⁹R¹⁰, —C(═O)C₁₋₈        alkyl or    -   —C(═O)CH(NH₂)CH₃;    -   R⁹ is hydrogen, C₁₋₈ alkyl or substituted C₁₋₈ alkyl;    -   R¹⁰ is hydrogen, C₁₋₈ alkyl or substituted C₁₋₈ alkyl;    -   R⁵ is hydrogen, C₁₋₈ alkyl, substituted C₁₋₈ alkyl, aryl,        substituted aryl, C₁₋₈ alkenyl, substituted C₁₋₈ alkenyl,        arylalkyl, substituted arylalkyl, alkylaryl, substituted        alkylaryl, aminoarylalkyl or substituted aminoarylalkyl;    -   R⁷ is —OH or halogen; and    -   R⁸ is hydrogen, halogen or C₁₋₈ alkyl,    -   provided that R² and R³ cannot both be hydrogen when R⁴ is a        halogen, Z is not present, R⁷ is —OH, R⁵, R⁶ and R⁸ are        hydrogen.

In another aspect, there is provided a compound as well as compositionsand methods using the same to prevent and/or treat a degenerativedisorder of the central nervous system in a patient at risk fordeveloping or diagnosed with the same which includes administering tothe patient in need thereof an effective amount of a compound defined byFormula II:

wherein:

-   -   R¹ is C(R²)(R³)(R⁴);    -   R² is hydrogen, —OH or halogen;    -   R³ is hydrogen, —OH, halogen or —CH₃;    -   R⁴ is halogen, —CH₃, phenyl, fluorophenyl, methylphenyl,        cyclohexylmethyl, wherein when R⁴ is a halogen, both R² and R³        cannot be hydrogen;    -   R³ and R⁴ may join with the carbon to which they are attached to        form a cycloalkyl ring, which may be optionally substituted with        one or more halogen atoms;    -   R⁶ is hydrogen, phenylalkyl or substituted phenylalkyl;    -   Z is optional, when present Z is —(CH₂)—, —C(═O) —, —S(═O)₂NH—,        —S(═O)₂—, —S(═O)₂—CH₃, C(═O)—NH—, —S(═O)₂NR⁹R¹⁰, —C(═S)—NH— or        —C(═O)₂—CH₃,    -   R⁹ is hydrogen or CH₃;    -   R¹⁰ is hydrogen or CH₃;    -   R⁵ is hydrogen or aminophenylalkyl;    -   R⁷ is —OH or halogen; and    -   R⁸ is hydrogen, halogen or —CH₃,    -   provided that R² and R³ cannot both be hydrogen when R⁴ is        halogen, Z is not present, R⁷ is —OH, R⁵, R⁶ and R⁸ are        hydrogen.

In yet another aspect, there is provided a compound as well ascompositions and methods using the same to prevent and/or treat adegenerative disorder of the central nervous system in a patient at riskfor developing or diagnosed with the same which includes administeringto the patient in need thereof an effective amount of a compound definedby Formula III:

wherein:

-   -   R¹ is C(R²)(R³)(R⁴);    -   R² is hydrogen, —OH or halogen;    -   R³ is hydrogen, —OH, halogen or —CH₃;    -   R⁴ is halogen, —CH₃, phenyl, fluorophenyl, methylphenyl,        cyclohexylmethyl, wherein when R⁴ is a halogen, both R² and R³        cannot be hydrogen;    -   R³ and R⁴ may join with the carbon to which they are attached to        form a cycloalkyl ring, which may be optionally substituted with        one or more halogen atoms;    -   R⁷ is —OH or halogen; and    -   R⁸ is hydrogen, halogen or —CH₃,    -   provided that R² and R³ cannot both be hydrogen when R⁴ is a        halogen,    -   R⁷ is —OH and R⁶ and R⁸ are hydrogen.

It is understood by a person of ordinary skill in the art that R², R³and R⁴ in aforementioned Formulas I, II, and III will not be selectedsuch that an unstable molecule will result.

In still another aspect, there is provided a compound as well ascompositions and methods using the same to prevent and/or treat adegenerative disorder of the central nervous system in a patient at riskfor developing or diagnosed with the same which includes administeringto the patient in need thereof an effective amount of a compoundselected from the following:

or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

In one embodiment, the compound is(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5S)-5-benzylpiperidine-3,4-diol, or a pharmaceutically acceptablesalt, solvate, or prodrug thereof. In one embodiment, the compound is(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol, or a pharmaceuticallyacceptable salt, solvate, or prodrug thereof. In one embodiment, thecompound is (3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol, or apharmaceutically acceptable salt, solvate, or prodrug thereof. In oneembodiment, the compound is (3R,4R,5S)-5-benzylpiperidine-3,4-diol, or apharmaceutically acceptable salt, solvate, or prodrug thereof.

In one embodiment, the degenerative disorder is a synucleinopathy. Inone embodiment, the degenerative disorder is characterized by Lewybodies. In one embodiment, the degenerative disorder is Parkinson'sdisease, dementia with Lewy bodies, multiple system atrophy orAlzheimer's disease. In one embodiment, the degenerative disorder isassociated with aggregation of at least one protein. In one embodiment,the degenerative disorder is associated with aggregation ofalpha-synuclein. In one embodiment, the degenerative disorder isassociated with aggregation of non-Abeta component. In one embodiment,the degenerative disorder is associated with accumulation of at leastone glycolipid. In one embodiment, the degenerative disorder isassociated with accumulation of at least one glycosphingolipid. In oneembodiment, the degenerative disorder is associated with accumulation ofglucocerebroside. In one embodiment, the degenerative disorder isassociated with a mutation in glucocerebrosidase. In one embodiment, themethod further comprises administering an effective amount of at leastone other therapeutic agent. In one embodiment, at least one othertherapeutic agent is levodopa, an anticholinergic, a catechol-O-methyltransferase inhibitor, a dopamine receptor agonist, a monoamine oxidaseinhibitor, a peripheral decarboxylase inhibitor, or an anti-inflammatoryagent.

The present invention also provides methods for preventing and/ortreating Parkinson's disease in a patient at risk for developing ordiagnosed with the same, which comprises administering to the patient inneed thereof an effective amount of any of the aforementioned compounds,or a pharmaceutically acceptable salt, solvate, or prodrug thereof, orany combination of two or more thereof.

In one embodiment, the method comprises administering the compound(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5S)-5-benzylpiperidine-3,4-diol, or a pharmaceutically acceptablesalt, solvate, or prodrug thereof. In one embodiment, the methodcomprises administering the compound(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol, or a pharmaceuticallyacceptable salt, solvate, or prodrug thereof. In one embodiment, themethod comprises administering the compound(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol, or a pharmaceuticallyacceptable salt, solvate, or prodrug thereof. In one embodiment, themethod comprises administering the compound(3R,4R,5S)-5-benzylpiperidine-3,4-diol, or a pharmaceutically acceptablesalt, solvate, or prodrug thereof.

In one embodiment, the method comprises administering an effectiveamount of at least one other therapeutic agent. In one embodiment, atleast one other therapeutic agent is levodopa, an anticholinergic, acatechol-β-methyl transferase inhibitor, a dopamine receptor agonist, amonoamine oxidase inhibitor, a peripheral decarboxylase inhibitor, or ananti-inflammatory agent.

The present invention also provides kits comprising:

-   -   a container having an effective amount of any of the compounds        of the present invention, alone or in combination; and    -   instructions for using the same to prevent and/or treat a        degenerative disorder of the central nervous system.        In one embodiment, the degenerative disorder of the central        nervous system is Parkinson's disease. In one embodiment, the        degenerative disorder of the central nervous system is        Alzheimer's disease.

DETAILED DESCRIPTION OF THE INVENTION

As used herein the following terms shall have the definitions set forthbelow.

As used herein, the phrase “degenerative disorder of the central nervoussystem” means any disorder associated with the premature degeneration ofany component of the central nervous system, such as neurons, myelinsheaths or axons. Such disorders include but are not limited tomulti-infarct dementia, Huntington's disease, Pick's disease,amyotrophic lateral sclerosis, Creutzfeldt-Jakob's disease, frontal-lobedegeneration, corticobasal degeneration, progressive supranuclear palsy,Parkinson's disease, dementia with Lewy bodies, multiple system atrophyor Alzheimer's disease.

As used herein the term “treating” means to ameliorate one or moresymptoms associated with the referenced disorder.

As used herein, the term “preventing” means to mitigate a symptom of thereferenced disorder.

As used herein the phrase “an effective amount” means an amounteffective to prevent and/or treat a patient at risk for developing ordiagnosed with the referenced disorder, and thus producing the desiredtherapeutic effect.

As used herein the term “patient” means a mammal (e.g., a human).

Listed below are chemical definitions of various terms used to describethis invention. These definitions apply to the terms as they are usedthroughout this specification, unless otherwise limited in specificinstances, either individually or as part of a larger group.

The term “alkyl” refers to straight or branched chain unsubstitutedhydrocarbon groups of 1 to 20 carbon atoms, preferably 1 to 8 carbonatoms, more preferably 1 to 6 carbon atoms. The expression “lower alkyl”refers to unsubstituted alkyl groups of 1 to 4 carbon atoms.

The term “substituted alkyl” refers to an alkyl group substituted by,for example, one to four substituents, such as, halo, hydroxy, alkoxy,oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino,aralkylamino, disubstituted amines in which the 2 amino substituents areselected from alkyl, aryl or aralkyl; alkanoylamino, aroylamino,aralkanoylamino, substituted alkanoylamino, substituted arylamino,substituted aralkanoylamino, thiol, alkylthio, arylthio, aralkylthio,alkylthiono, arylthiono, aralkylthiono, alkylsulfonyl, arylsulfonyl,aralkylsulfonyl, sulfonamido, e.g. SO₂NH₂, substituted sulfonamido,nitro, cyano, carboxy, carbamyl, e.g. CONH₂, substituted carbamyl e.g.CONHalkyl, CONHaryl, CONHaralkyl or cases where there are twosubstituents on the nitrogen selected from alkyl, aryl or aralkyl;alkoxycarbonyl, aryl, substituted aryl, guanidino and heterocyclos, suchas, indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl,pyrimidyl and the like. Where noted above where the substituent isfurther substituted it will be with alkyl, alkoxy, aryl or aralkyl.

The term “halogen” or “halo” refers to fluorine, chlorine, bromine andiodine.

The term “aryl” refers to monocyclic or bicyclic aromatic hydrocarbongroups having 6 to 12 carbon atoms in the ring portion, such as phenyl,naphthyl, biphenyl and diphenyl groups, each of which may besubstituted.

The term “aralkyl” refers to an aryl group bonded directly through analkyl group, such as benzyl. Similarly, the term “alkylaryl” refers toan alkyl group bonded directly through an aryl group, such asmethylbenzyl.

The term “substituted aryl” refers to an aryl group substituted by, forexample, one to four substituents such as alkyl, substituted alkyl,halo, trifluoromethoxy, trifluoromethyl, hydroxy, alkoxy, alkanoyl,alkanoyloxy, amino, alkylamino, aralkylamino, dialkylamino,alkanoylamino, thiol, alkylthio, ureido, nitro, cyano, carboxy,carboxyalkyl, carbamyl, alkoxycarbonyl, alkylthiono, arylthiono,arylsulfonylamine, sulfonic acid, alkysulfonyl, sulfonamido, aryloxy andthe like. The substituent may be further substituted by hydroxy, alkyl,alkoxy, aryl, substituted aryl, substituted alkyl or aralkyl.

The term “heteroaryl” refers to an optionally substituted, aromaticgroup for example, which is a 4 to 7 membered monocyclic, 7 to 11membered bicyclic, or 10 to 15 membered tricyclic ring system, which hasat least one heteroatom and at least one carbon atom-containing ring,for example, pyridine, tetrazole, indazole.

The term “alkenyl” refers to straight or branched chain hydrocarbongroups of 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, andmost preferably 2 to 8 carbon atoms, having one to four double bonds.

The term “substituted alkenyl” refers to an alkenyl group substitutedby, for example, one to two substituents, such as, halo, hydroxy,alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino,alkanoylamino, thiol, alkylthio, alkylthiono, alkylsulfonyl,sulfonamido, nitro, cyano, carboxy, carbamyl, substituted carbamyl,guanidino, indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl,pyridyl, pyrimidyl and the like.

The term “alkynyl” refers to straight or branched chain hydrocarbongroups of 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, andmost preferably 2 to 8 carbon atoms, having one to four triple bonds.

The term “substituted alkynyl” refers to an alkynyl group substitutedby, for example, a substituent, such as, halo, hydroxy, alkoxy,alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino, alkanoylamino,thiol, alkylthio, alkylthiono, alkylsulfonyl, sulfonamido, nitro, cyano,carboxy, carbamyl, substituted carbamyl, guanidino and heterocyclo, e.g.imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyland the like.

The term “cycloalkyl” refers to an optionally substituted, saturatedcyclic hydrocarbon ring systems, preferably containing 1 to 3 rings and3 to 7 carbons per ring which may be further fused with an unsaturatedC3-C7 carbocylic ring. Exemplary groups include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cycloctyl, cyclodecyl,cyclododecyl, and adamantyl. Exemplary substituents include one or morealkyl groups as described above, or one or more groups described aboveas alkyl substituents.

The terms “heterocycle”, “heterocyclic” and “heterocyclo” refer to anoptionally substituted, fully saturated or unsaturated, aromatic ornonaromatic cyclic group, for example, which is a 4 to 7 memberedmonocyclic, 7 to 11 membered bicyclic, or 10 to 15 membered tricyclicring system, which has at least one heteroatom in at least one carbonatom-containing ring. Each ring of the heterocyclic group containing aheteroatom may have 1, 2 or 3 heteroatoms selected from nitrogen atoms,oxygen atoms and sulfur atoms, where the nitrogen and sulfur heteroatomsmay also optionally be oxidized and the nitrogen heteroatoms may alsooptionally be quaternized. The heterocyclic group may be attached at anyheteroatom or carbon atom.

Exemplary monocyclic heterocyclic groups include pyrrolidinyl, pyrrolyl,indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl,imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl, isoxazolyl,thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl,furyl, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl,2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxazepinyl,azepinyl, 4-piperidonyl, pyridyl, N-oxo-pyridyl, pyrazinyl, pyrimidinyl,pyridazinyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl,thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane andtetrahydro-1,1-dioxothienyl, dioxanyl, isothiazolidinyl, thietanyl,thiiranyl, triazinyl, and triazolyl, and the like.

Exemplary bicyclic hetrocyclic groups include2,3-dihydro-2-oxo-1H-indolyl, benzothiazolyl, benzoxazolyl,benzothienyl, quinuclidinyl, quinolinyl, quinolinyl-N-oxide,tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl,indolizinyl, benzofuryl, chromonyl, coumarinyl, cinnolinyl,quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl (such asfuro[2,3-c]pyridinyl, furo[3,1-b]pyridinyli or furo[2,3-b]pyridinyl),dihydroisoindolyl, dihydroquinazolinyl (such as3,4-dihydro-4-oxo-quinazolinyl), benzisothiazolyl, benzisoxazolyl,benzodiazinyl, benzofurazanyl, benzothiopyranyl, benzotriazolyl,benzpyrazolyl, dihydrobenzofuryl, dihydrobenzothienyl,dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone,dihydrobenzopyranyl, indolinyl, isochromanyl, isoindolinyl,naphthyridinyl, phthalazinyl, piperonyl, purinyl, pyridopyridyl,quinazolinyl, tetrahydroquinolinyl, thienofuryl, thienopyridyl,thienothienyl, and the like.

Exemplary substituents include one or more alkyl or aralkyl groups asdescribed above or one or more groups described above as alkylsubstituents.

Also included are smaller heterocyclos, such as, epoxides andaziridines.

The term “heteroatoms” shall include oxygen, sulfur and nitrogen.

Parkinson's disease may be diagnosed in patients according to the UnitedKingdom Parkinson's Disease Society brain-bank clinical diagnosticcriteria (see, Hughes et al., Accuracy of clinical diagnosis ofidiopathic Parkinson's disease: a clinico-pathological study of 100cases. J Neurol Neurosurg Psychiatry 1992; 55:181-184) and/or thecriteria described by Gelb et al., Diagnostic Criteria for Parkinson'sDisease. Arch Neurol. 1999; 56(1):33-39. Likewise, the severity ofParkinson's disease may be ascertained using the Unified Parkinson'sDisease Rating Scale. See, e.g., Fahn and Elton, Members of the UnifiedParkinson's Disease Rating Scale Development Committee. UnifiedParkinson's Disease Rating Scale. In: Fahn et al., Recent developmentsin Parkinson's disease. New York: Macmillan, 1987: 153-163.

Alzheimer's disease may be diagnosed in patients according to thecriteria for dementia of the Alzheimer's type of the Diagnostic andStatistical Manual of Mental Disorders, 4^(th) ed.: DSM-IV. Washington,D.C.: American Psychiatric Association, 1994. Likewise, the criteria forprobable Alzheimer's disease may be ascertained based on criteria of theNational Institute of Neurological and Communicative Disorders andStroke and the Alzheimer's Disease and Related Disorders Association.See also, McKhann et al., Clinical diagnosis of Alzheimer's disease:report of the NINCDS-ADRDA work group under the auspices of Departmentof Health and Human Services Task Force on Alzheimer's Disease.Neurology 1984; 34:939-944.

Multiple system atrophy (MSA) is characterized by glial cytoplasmicinclusion bodies (also known as Papp-Lantos bodies) in the movement,balance and automatic control centers of the brain. The most commonfirst sign of MSA is the appearance of an “akinetic-rigid syndrome”(i.e., slowness of initiation of movement resembling Parkinson'sdisease) found in 62% at first presentation. Other common signs at onsetinclude problems with balance (found in 22%), followed by genito-urinaryproblems (9%). For men, the first sign can be erectile dysfunction(unable to achieve or sustain an erection). Both men and women oftenexperience problems with their bladders including urgency, frequency,incomplete bladder emptying or an inability to pass urine (retention).About 1 in 5 MSA patients will suffer a fall in their first year ofdisease. As the disease progresses three groups of symptoms predominate.These are: (i) parkinsonism (slow, stiff movement, writing becomes smalland spidery); (ii) cerebellar dysfunction (difficulty coordinatingmovement and balance); and (iii) autonomic dysfunction (impairedautomatic body functions) including: postural or orthostatichypotension, resulting in dizziness or fainting upon standing up,urinary incontinence, impotence; constipation; dry mouth and skin;trouble regulating body temperature due to abnormal sweating; abnormalbreathing during sleep. Notably, not all of these symptoms areexperienced by all patients.

Dementia with Lewy bodies (DLB) is one of the most common types ofprogressive dementia. The central feature of DLB is progressivecognitive decline, combined with three additional defining features: (1)pronounced “fluctuations” in alertness and attention, such as frequentdrowsiness, lethargy, lengthy periods of time spent staring into space,or disorganized speech; (2) recurrent visual hallucinations, and (3)parkinsonian motor symptoms, such as rigidity and the loss ofspontaneous movement. People may also suffer from depression. Thesymptoms of DLB are caused by the build-up of Lewy bodies—accumulatedbits of alpha-synuclein protein—inside the nuclei of neurons in areas ofthe brain that control particular aspects of memory and motor control.Researchers don't know exactly why alpha-synuclein accumulates into Lewybodies or how Lewy bodies cause the symptoms of DLB, but they do knowthat alpha-synuclein accumulation is also linked to Parkinson's disease,multiple system atrophy, and several other disorders, which are referredto as the “synucleinopathies.” The similarity of symptoms between DLBand Parkinson's disease, and between DLB and Alzheimer's disease, canoften make it difficult for a doctor to make a definitive diagnosis. Inaddition, Lewy bodies are often also found in the brains of people withParkinson's and Alzheimer's diseases. These findings suggest that eitherDLB is related to these other causes of dementia or that an individualcan have both diseases at the same time. DLB usually occurssporadically, in people with no known family history of the disease.However, rare familial cases have occasionally been reported.

Compounds

Novel compounds of the present invention are provided below:

Chemical Process

Compositions of the present invention can be made in accordance of oneor more of the following schemes.

((2S,3S,4aR,8R,8aR)-2,3-Dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridin-8-yl)methanolHydrochloride(2)

A solution of 1 (20.0 g, 55.0 mmol) in MeOH (500 mL) was combined withPd(OH)₂ (4-6 g) and ammonium formate (14 g, 220 mmol) and the mixturewas heated at 50-55° C. Additional amounts (3×100.0 mmol) of ammoniumformate were added over the next 8 hrs. After the final addition, thereaction mixture was further stirred and heated an additional 16 hrs at50-55° C. The catalyst was removed by filtration and the filtrate wasevaporated in vacuo. The crude product was dissolved in acetone (150mL), filtered, and HCl in 2-PrOH was added. After seeding and thencooling in an ice bath, the product was collected as a white crystallinesolid (11.0 g, 71%). ¹H NMR (DMSO-d₆) 9.45 (s, 2H), 4.80 (t, 1H, ex),3.85 (m, 1H), 3.0-3.75 (m, 11H), 2.8 (q, 2H), 1.95 (m, 1H), 1.2 (2, 6H).

((2S,3S,4aR,8R,8aR)-6-Benzyl-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridin-8-yl)methanol(3)

To a solution of 2 (14.85 g, 50.0 mmol) in DMF (200 mL) was added K2CO3(17.25 g, 125 mmol) and the mixture was stirred at 40° C. for about 4hrs. At this point, BnCl (5.7 mL, 50.0 mmol) was added in one portionand the reaction was stirred at 40° C. overnight. The solvent wasevaporated in vacuo and the residue was suspended in water (600 mL) andHCl was added to dissolve the residue. The solution was washed with Et2Oand then basified with Na2CO3. The solution was extracted with EtOAc(2×) and the combined extracts were washed with water and then brine andthen dried over MgSO4. The solution was filtered and the filtrateevaporated in vacuo to give the title compound (17.2 g, >95%) as acolorless to very pale yellow viscous oil which was used without furtherpurification. 1H NMR (CDCl3) 7.3 (m, 5H), 3.6-3.8 (m, 2H), 3.5 (s, 3H),3.4 (t, 1H), 3.26 (s, 3H), 3.268 (s, 3H), 2.9 (m, 2H), 2.2 (br s, 1H),2.05 (m, 1H), 1.85 (t, 1H), 1.28 (s, 3H), 1.26 (s, 3H).

((2S,3S,4aR,8R,8aR)-6-Benzyl-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridin-8-yl)carboxaldehyde(General Procedure A) (4)

To a solution of DMSO (7.3 g, 96.9 mmol) in CH2Cl2 (150 mL) cooled to−78° C. was added a solution of oxalyl chloride (6.1 mL, 72.8 mmol) inCH2Cl2 dropwise. After the addition was complete the reaction mixturewas stirred for an additional 30 min at which point a solution of 3(17.0 g, 48.4 mmol) in CH2Cl2 was added dropwise. After addition wascomplete, the reaction was stirred for 1 hr at −78° C. and thendiisopropylethylamine (34.4 mL, 193 mmol) was added dropwise. After thisaddition was complete, the cooling bath was removed and the reactionmixture was allowed to warm to 0° C. when saturated NaHCO3 was added.The mixture was diluted with some additional CH2Cl2 and then the organiclayer was separated and dried over MgSO4. After filtering, the solventwas evaporated in vacuo and the crude product was purified by silica gelchromatography (Hex/EtOAc) to give the title compound (12.7 g, 75%) as aviscous oil. 1H NMR (CDCl3) 9.73 (s, 1H), 7.2 (m, 5H), 3.75 (m, 2H), 3.5(q, 2H), 3.2 (2 s, 6H), 2.7-3.0 (m, 3H), 2.05 (m, 2H), 1.25 (2 s, 6H).

((2S,3S,4aR,8S,8aR)-6-Benzyl-8,8-difluoromethyl-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridineHydrochloride (General Procedure B) (5)

To a solution of DAST (1.4 mL, 10.3 mmol) in CH₂Cl₂ (50 mL) cooled to−15° C. was added a solution of 4 (2.4 g, 6.9 mmol) dropwise. After 10minutes, the ice bath was removed and the reaction was stirred at roomtemperature overnight. At this point the reaction mixture was againcooled in an ice bath and the reaction was quenched by addition ofsaturated NaHCO3 (dropwise at first since this does produce a slightexotherm). The organic layer was separated and dried over Na2SO4,filtered and the solvent was evaporated in vacuo to give a yellow oil.The residue was purified by chromatography on silica gel (Hex/EtOAc) togive the title compound (1.6 g, 62%) as a colorless oil. 1H NMR (CDCl3)7.2 (m, 5H), 6.0 (dt, 1H), 3.75 (m, 1H), 3.55 (m, 3H), 3.2 (2s, 6H),2.95 (m, 1H), 2.85 (m, 1H), 2.3 (m, 2H), 1.5 (br s, 1H), 1.2 (2s, 6H).

(3R,4R,5S)-5-(Difluoromethyl)piperidine 3,4-diol Hydrochloride (GeneralProcedure C) (6)

Compound 5 (1.6 g, 4.3 mmol) was heated at reflux in a mixture ofEtOH/H2O/HCl (40 mL/40 mL/5 mL) and the reaction monitored by HPLC untilthe starting material could no longer be detected. The solvent wasevaporated in vacuo and then co-evaporated 2× with EtOH. The residue wasdissolved in MeOH and hydrogenated over Pd(OH)2. When complete, thecatalyst was removed by filtration and the filtrate evaporated in vacuo.The residue was recrystallized from EtOH (50 mL) to the title compound(0.55 g, 66%) as a white solid (mp 168-170° C.). 1H NMR (D2O) 6.15 (dt,1H), 4.3-4.8 (m, 2H), 3.0 (t, 1H), 2.85 (t, 1H), 2.3 (m, 1H).

(R) and(S)-1-((2S,3S,4aR,8R,8aR)-6-Benzyl-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridin-8-yl)ethanolGeneral Procedure D(15/16)

To a solution of 4 (7.0 g, 20.0 mmol) in dry THF (100 mL) was addedMeMgBr (20.0 mL, 1.4 M in 3:1 THF/toluene) and the reaction was stirredovernight at room temperature. The reaction was quenched with saturatedNH4Cl and the mixture was extracted with EtOAc (2×). The combinedextracts were washed with brine, dried over Na2SO4 and the filtrate wasevaporated in vacuo. The residue was purified by silica gelchromatography (hexane/2-PrOH) to give the major isomer (15) (1.6 g,24.6%). 1H NMR (CDCl3). 7.3 (m, 5H), 4.15 (m, 1H), 3.5-3.9 (m, 3H), 3.3(2 s, 6H), 2.85 (m, 2H), 2.0 (2 m, 4H), 1.3 (2 s, 6H), 1.2 (d, 3H). Theminor isomer (16) was also isolated (0.55 g, 7.5%) 7.3 (m, 5H), 3.75 (m,2H), 3.5 (m, 2H), 3.2 (2 s, 6H), 2.8 (m, 2H), 2.0 (t, 1H), 1.75 (m, 2H),1.2 (2 s, 6H), 1.0 (d, 3H).

(3R,4R,5R)-5((R)-1-Hydroxyethyl)piperidine 3,4-diol (17)

Compound 15 (0.55 g, 1.5 mmol) was stirred in a mixture of 9/1 TFA:H2O(20 mL) until the starting material could no longer be detected by HPLC.The volatiles were removed and the residue was co-evaporated 2-3× withEtOH and then dissolved in EtOH and treated with solid K2CO3. Afterfiltering the solid, the filtrate was evaporated in vacuo, and theresidue was converted to an HCl salt and hydrogenated over Pd(OH)2. Thecatalyst was filtered and the filtrate evaporated in vacuo. The crudeproduct was purified using an ion exchange resin (Dowex 50WX8-200)eluting with 0.1 N NH4OH. The appropriate fractions were combined andlyophilized to give the title compound (0.12 g, 50%). 1H NMR (D2O) 4.2(q, 1H), 3.65 (m, 1H), 3.45 (m, 3H), 2.8 (m, 2H), 1.65 (m, 1H), 1.15 (d,3H).

(3R,4R,5R)-5((S)-1-Hydroxyethyl)piperidine 3,4-diol (10)

Compound 16 (0.34 g, 0.93 mmol) was deprotected as described above togive the title compound (0.11 g, 75%). 1H NMR (D2O) 4.15 (m, 2H), 3.5(m, 1H), 3.35 (t, 1H), 3.15 (m, 2H), 1.8 (m, 1H), 1.1 (d, 3H).

((2S,3S,4aR,8R,8aR)-6-Benzyl-8(S)-(1-fluoroethyl)-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridine(11)

Compound 15 (1.8 g, 5.0 mmol) was fluorinated using General Procedure B.Silica gel chromatography (Hex/EtOAc) gave the title compound (0.42 g,23%). 1H NMR (CDCl3) 7.25 (m, 5H), 4.7-4.9 (dq, 1H), 3.75 (m, 2H), 3.4(m, 2H), 3.2 (2 s, 6H), 2.8 (m, 2H), 2.0 (m, 3H), 1.35 (dd, 3H), 1.2 (2s, 6H).

(3R,4R,5R)-5((S)-1-Fluoroethyl)piperidine 3,4-diol Hydrochloride (13)

Compound II (0.42 g, 1.14 mmol) was deprotected as described in GeneralProcedure C. After catalyst was removed, the filtrate was evaporated invacuo and then co-evaporated with EtOH (2×). The resulting residue wastriturated with acetone to give the title compound (0.20 g, 88%) as awhite solid. 1H NMR (DMSO-d6) 9.0 (br s, 2H), 5.6 (d, 1H, ex), 5.4 (d,1H, ex), 5.0-5.2 (dq, 1H), 3.55 (m, 1H), 3.2 (m, 2H), 2.9 (t, 1H), 2.7(t, 1H), 2.2 (m, 1H), 1.3 (dd, 3H).

((2S,3S,4aR,8R,8aR)-6-Benzyl-8(R)-(1-fluoroethyl)-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridine(12)

Compound 16 (0.55 g, 1.5 mmol) was fluorinated using General Procedure Bto give the title compound (0.22 g, 40%). 1H NMR (CDCl3) 7.3 (m, 5H),5.0 (dq, 1H), 3.8 (m, 1H), 3.5-3.75 (m, 3H), 3.3 (2 s, 6H), 3.0 (d, 1H),2.9 (m, 1H), 2.1 (m, 2H), 1.85 (m, 1H), 1.3 (2 s, 6H).

(3R,4R,5R)-5((R)-(1-Fluoroethyl)piperidine 3,4-diol Hydrochloride (14)

Compound 12 (0.22 g, 0.6 mmol) was deprotected as described in GeneralProcedure C. After catalyst was removed, the filtrate was evaporated invacuo and then co-evaporated with EtOH (2×). The resulting residue wastriturated with acetone to give the title compound (0.08 g, 67%) as awhite solid. 1H NMR (D2O) 5.1 (dq, 1H), 3.5 (m, 4H), 2.8 (m, 2H), 1.8(m, 1H), 1.3 (dd, 3H).

((2S,3S,4aR,8R,8aR)-2,3-Dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridin-8-yl)methanolHydrochloride(2)

A solution of 1 (20.0 g, 55.0 mmol) in MeOH (500 mL) was combined withPd(OH)2 (4-6 g) and ammonium formate (14 g, 220 mmol) and the mixturewas heated at 50-55° C. Additional amounts (3×100.0 mmol) of ammoniumformate were added over the next 8 hrs. After the final addition, thereaction mixture was further stirred and heated an additional 16 hrs at50-55° C. The catalyst was removed by filtration and the filtrate wasevaporated in vacuo. The crude product was dissolved in acetone (150mL), filtered, and HCl in 2-PrOH was added. After seeding and thencooling in an ice bath, the product was collected as a white crystallinesolid (11.0 g, 71%). ¹H NMR (DMSO-d₅) 9.45 (s, 2H), 4.80 (t, 1H, ex),3.85 (m, 1H), 3.0-3.75 (m, 11H), 2.8 (q, 2H), 1.95 (m, 1H), 1.2 (2, 6H).

((2S,3S,4aR,8R,8aR)-6-Benzyl-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridin-8-yl)methanol(3)

To a solution of 2 (14.85 g, 50.0 mmol) in DMF (200 mL) was added K₂CO₃(17.25 g, 125 mmol) and the mixture was stirred at 40° C. for about 4hrs. At this point, BnCl (5.7 mL, 50.0 mmol) was added in one portionand the reaction was stirred at 40° C. overnight. The solvent wasevaporated in vacuo and the residue was suspended in water (600 mL) andHCl was added to dissolve the residue. The solution was washed with Et₂Oand then basified with Na₂CO₃. The solution was extracted with EtOAc(2×) and the combined extracts were washed with water and then brine andthen dried over MgSO₄. The solution was filtered and the filtrateevaporated in vacuo to give the title compound (17.2 g, >95%) as acolorless to very pale yellow viscous oil which was used without furtherpurification. ¹H NMR (CDCl₃) 7.3 (m, 5H), 3.6-3.8 (m, 2H), 3.5 (s, 3H),3.4 (t, 1H), 3.26 (s, 3H), 3.268 (s, 3H), 2.9 (m, 2H), 2.2 (br s, 1H),2.05 (m, 1H), 1.85 (t, 1H), 1.28 (s, 3H), 1.26 (s, 3H).

((2S,3S,4aR,8R,8aR)-6-Benzyl-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridin-8-yl)carboxaldehyde(General Procedure A) (4)

To a solution of DMSO (7.3 g, 96.9 mmol) in CH₂Cl₂ (150 mL) cooled to−78° C. was added a solution of oxalyl chloride (6.1 mL, 72.8 mmol) inCH₂Cl₂ dropwise. After the addition was complete the reaction mixturewas stirred for an additional 30 min at which point a solution of 3(17.0 g, 48.4 mmol) in CH₂Cl₂ was added dropwise. After addition wascomplete, the reaction was stirred for 1 hr at −78° C. and thendiisopropylethylamine (34.4 mL, 193 mmol) was added dropwise. After thisaddition was complete, the cooling bath was removed and the reactionmixture was allowed to warm to 0° C. when saturated NaHCO₃ was added.The mixture was diluted with some additional CH₂Cl₂ and then the organiclayer was separated and dried over MgSO₄. After filtering, the solventwas evaporated in vacuo and the crude product was purified by silica gelchromatography (Hex/EtOAc) to give the title compound (12.7 g, 75%) as aviscous oil. ¹H NMR (CDCl₃) 9.73 (s, 1H), 7.2 (m, 5H), 3.75 (m, 2H), 3.5(q, 2H), 3.2 (2 s, 6H), 2.7-3.0 (m, 3H), 2.05 (m, 2H), 1.25 (2 s, 6H).

((2S,3S,4aR,8S,8aR)-6-Benzyl-8,8-difluoromethyl-2,3-dimethoxy-2,3-dimethyloctahydro-[1,4]dioxino[2,3-c]pyridineHydrochloride (General Procedure B) (5)

To a solution of DAST (1.4 mL, 10.3 mmol) in CH2Cl2 (50 mL) cooled to−15° C. was added a solution of 4 (2.4 g, 6.9 mmol) dropwise. After 10minutes, the ice bath was removed and the reaction was stirred at roomtemperature overnight. At this point the reaction mixture was againcooled in an ice bath and the reaction was quenched by addition ofsaturated NaHCO₃ (dropwise at first since this does produce a slightexotherm). The organic layer was separated and dried over Na₂SO₄,filtered and the solvent was evaporated in vacuo to give a yellow oil.The residue was purified by chromatography on silica gel (Hex/EtOAc) togive the title compound (1.6 g, 62%) as a colorless oil. ¹H NMR (CDCl3)7.2 (m, 5H), 6.0 (dt, 1H), 3.75 (m, 1H), 3.55 (m, 3H), 3.2 (2 s, 6H),2.95 (m, 1H), 2.85 (m, 1H), 2.3 (m, 2H), 1.5 (br s, 1H), 1.2 (2 s, 6H).

(3R,4R,5S)-5-(Difluoromethyl)piperidine 3,4-diol Hydrochloride (GeneralProcedure C) (6)

Compound 5 (1.6 g, 4.3 mmol) was heated at reflux in a mixture ofEtOH/H₂O/HCl (40 mL/40 mL/5 mL) and the reaction monitored by HPLC untilthe starting material could no longer be detected. The solvent wasevaporated in vacuo and then co-evaporated 2× with EtOH. The residue wasdissolved in MeOH and hydrogenated over Pd(OH)₂. When complete, thecatalyst was removed by filtration and the filtrate evaporated in vacuo.The residue was recrystallized from EtOH (50 mL) to the title compound(0.55 g, 66%) as a white solid (mp 168-170° C.). ¹H NMR (D₂O) 6.15 (dt,1H), 4.3-4.8 (m, 2H), 3.0 (t, 1H), 2.85 (t, 1H), 2.3 (m, 1H).

(3R,4R,5S)-1.Butyl-5-(difluoromethyl)piperidine 3,4-diol (GeneralProcedure D) (7a; R⁵=Bu)

A mixture of 6 (0.30 g, 1.4 mmol), K₂CO₃ (0.48 g, 3.5 mmol) and BuBr(0.20 g, 1.4 mmol) was combined in DMF (10 mL) and heated overnight at60° C. The solvent was evaporated in vacuo and the residue was dissolvedin EtOAc, washed with water and then brine and dried over Na₂SO₄. Afterfiltration, the filtrate was evaporated in vacuo to give the crudeproduct which was purified by chromatography (CH₂Cl₂/(9:1) MeOH/NH₄OH)to give the title compound (0.25 g, 80%) as a colorless sirup. MH⁺=224.¹H NMR (DMSO-d₆) 6.2 (t, 1H, J=57 Hz), 5.13 (d, 1H, ex), 4.91 (d, 1H,ex), 3.3 (m, 1H), 3.1 (m, 1H), 2.9 (m, 2H), 2.3 (m, 2H), 1.95 (m, 2H),1.75 (t, 1H), 1.2-1.5 (2 m, 4H), 0.9 (t, 3H).

(3R,4R,5S)-1.Allyl-5-(difluoromethyl)piperidine 3,4-diol (7b; R⁵=allyl)

Following General Procedure D using allyl bromide (0.17 g, 1.4 mmol) thetile compound was obtained as a white solid (0.22 g, 76%). MN⁺=208. ¹HNMR (DMSO-d₆) 6.2 (t, 1H, J=57 Hz), 5.8 (m, 1H), 5.2 (m, 3H), 4.92 (d,1H), 3.3 (m, 1H), 3.1 (1H), 2.95 (d, 2H), 2.85 (d, 2H), 1.9 (br m, 2H),1.75 (t, 1H).

(3R,4R,5S)-5-(Difluoromethyl)-1-(4-fluorobenzyl)piperidine 3,4-diol (7c;R⁵=4-fluorobenzyl).

Following General Procedure D except that reaction was run at roomtemperature and using 4-fluorobenzyl bromide (0.26 g, 1.4 mmol) the tilecompound was obtained as a white solid (0.22 g, 56%). MH⁺=276. ¹H NMR(DMSO-d₆) 7.4 (m, 2H), 7.15 (m, 2H), 6.2 (t, 1H, J=57 Hz), 5.2 (d, 1H,ex), 4.9 (d, 1H, ex), 3.5 (q, 2H), 3.3 (m, 1H), 3.1 (m, 1H), 2.8 (m,2H), 2.0 (m, 2H), 1.8 (t, 1H).

(3R,4R,5S)-5-(Difluoromethyl)-1-(4-methylbenzyl)piperidine 3,4-diol (7d;R⁵=4-methylbenzyl)

Following General Procedure D except that reaction was run at roomtemperature and using 4-methylbenzyl bromide (0.26 g, 1.4 mmol) the tilecompound was obtained as a white solid (0.30, 81%). MH⁺=272. ¹H NMR(DMSO-d₆) 7.2 (m, 4H), 6.2 (t, 1H, J=57 Hz), 5.2 (d, 1H, ex), 4.9 (d,1H, ex), 3.5 (q, 2H), 3.3 (1H), 3.05 (m, 1H), 2.8 (m, 2H), 2.5 (s, 3H),1.95 (m, 2H), 1.8 (t, 1H).

(3R,4R,5S)-5-(Difluoromethyl)-1-(4-methoxylbenzyl)piperidine 3,4-diol(7e; R⁵=4-methoxylbenzyl)

Following General Procedure D except that reaction was run at roomtemperature and using 4-methoxylbenzyl chloride (0.26 g, 1.4 mmol) thetile compound was obtained as a colorless sirup (0.19 g, 49%). MH⁺=288.¹H NMR (DMSO-d₆) 7.3 (m, 1H), 6.85 (m, 3H) 6.2 (t, 1H, J=57 Hz), 5.2 (d,1H, ex), 4.9 (d, 1H, ex), 3.75 (s, 3H), 3.5 (q, 2H), 3.4 (m, 1H), 3.1(m, 1H), 2.85 (m, 2H), 1.95 (m, 2H), 1.8 (t, 1H).

1-((3S,4R,5R)-3-(Difluoromethyl)-4,5-dihydroxypiperidine-1-yl)pentane-1-one(8a; Z═CO; R⁵=butyl)

Following General Procedure D, except that the reaction was run at roomtemperature and using pentanoyl chloride (0.17 g, 1.4 mmol), the titlecompound was obtained as a white solid (0.26 g, 71%). MH⁺=252. ¹H NMR(DMSO-d₆) 5.9-6.5 (dt, 1H), 5.35 (m, 1H, ex), 5.25 (m, 1H), ex), 4.2(dd, 1H), 3.75 (dd, 1H), 3.35 (m, 2H), 3.1 (m, 1H), 2.85 (m, 1H), 2.3(t, 2H), 1.9 br m, 1H), 1.4 (m, 2H), 1.25 (m, 2H), 0.85 (t, 3H).

(3R,4R,5S)-5-(Difluoromethyl)-1-(methanesulfonyl)piperidine 3,4-diol(8b; Z═SO2; R⁵=Me)

Following General Procedure D except that the reaction was run at roomtemperature and using methanesulfonyl chloride (0.16 g, 1.4 mmol), thetitle compound was obtained as a white solid (0.17 g, 51%). 1H NMR(DMSO-d₆) 6.2 (t, 1H, J=53 Hz), 5.43 (d, 1H, ex), 5.38 (d, 1H, ex),3.2-3.7 (m, 4H), 2.95 (s, 3H), 2.85 (m, 1H), 2.7 (t, 1H), 2.1 (br s,1H). (3R,4R,5S)-5-(Difluoromethyl)-1-tosylpiperidine 3,4-diol (8b;Z═SO2; R⁵=Ph) Following General Procedure D except that the reaction wasrun at room temperature and using toluenesulfonyl chloride (0.26, 1.4mmol), the title compound was obtained as a white solid (0.35 g, 67%).¹H NMR (DMSO-d₆) 7.6 (d, 2H), 7.45 (d, 2H), 6.25 (t, 1H, J=53 Hz), 5.4(2d, 2H, ex), 3.3-3.55 (m, 4H), 3.2 (m, 1H), 2.5 (m, 3H), 2.4 (t, 1H),2.1 (m, 1H).

(3S,4R,5R)-3-(Difluoromethyl)-4,5-dihydroxy-N-propylpiperidine-1-carboxamide(General Procedure E) (9a; X═O; R⁵=propyl)

To a solution of 6 (free base) (0.29 g, 1.2 mmol) in dry DMF (5 mL), wasadded propyl isocyanate (0.10 g, 1.2 mmol) and the reaction was stirredat room temperature overnight. The solvent was evaporated in vacuo andthe residue was purified by chromatography (CH₂Cl₂/MeOH) to give thetitle compound as a white solid (0.14 g, 48%). MH⁺=253. ¹H NMR (DMSO-d₆)6.7 (t, 1H), 6.22 (t, 1H, J=53 Hz), 5.25 (d, 1H, ex), 5.15 (d, 1H, ex),4.05 (d, 1H), 3.9 (d, 1H), 3.3 (m, 2H), 3.0 (q, 2H), 2.5 (m, 1H), 1.8(br d, 1H), 1.4 (m, 2H), 0.85 (t, 3H).

(3S,4R,5R)-3-(Difluoromethyl)-4,5-dihydroxy-N-phenylpiperidine-1-carboxamide(9b; X═O; R⁵=phenyl)

Following General Procedure E and using phenyl isocyanate (0.14 g, 1.2mmol) the title compound was obtained as a white solid (0.21 g, 62%).MH⁺=287. ¹H NMR (DMSO-d6) 8.7 (s, 1H), 7.45 (d, 2H), 7.3 (t, 2H), 6.95(t, 1H), 6.3 (t, 1H, J=53 Hz), 5.35 (d, 1H), 5.25 (d, 1H), 4.1 (t, 2H),3.3 (m, 2H), 2.85 (t, 1H), 2.75 (t, 1H), 1.95 (br d, 1H).

(3S,4R,5R)-3-(Difluoromethyl)-4,5-dihydroxy-N-butylpiperidine-1-carboxamide(9c; X═O; R⁵=butyl)

Following General Procedure E and using butyl isocyanate (0.12 g, 1.2mmol) the title compound was obtained as a white solid (0.24 g, 76%).MH⁺=267. ¹H NMR (DMSO-d₆) 6.6 (t, 1H), 6.2 (t, 1H, J=53 Hz), 5.25 (d,1H), 5.1 (d, 1H), 4.05 (d, 1H), 3.9 (d, 1H), 3.35 (m, 2H), 3.05 (q, 2H),2.65 (t, 1H), 2.45 (m, 1H), 1.8 (br d, 1H), 1.2-1.4 (2 m, 4H), 0.85 (t,3H).

(3S,4R,5R)-3-(Difluoromethyl)-4,5-dihydroxy-N-butylpiperidine-1-carbthioamide(9d; X═S; R⁵=butyl)

Following General Procedure E and using butyl isothiocyanate (0.14 g,1.2 mmol) the title compound was obtained as a colorless sirup (0.21 g,63%). MH⁺=283. ¹H NMR (DMSO-d₆) 7.85 (t, 1H), 6.25 (t, 1H), 5.35 (2d,2H), 4.8 (d, 1H), 4.45 (d, 1H), 3.45 (m, 2H), 3.25 (m, 1H), 3.05 (t,1H), 2.8 (t, 1H), 1.85 (br d, 1H), 1.4 (m, 2H), 1.35 (m, 2H), 1.1 (m,1H), 0.95 (t, 3H).

(3S,4R,5R)-3-(Difluoromethyl)-4,5-dihydroxy-N-phenylpiperidine-1-carbthioamide(9e; X═S; R⁵=phenyl)

Following General Procedure E and using phenyl isothiocyanate (0.16 g,1.2 mmol) the title compound was obtained as a white solid (0.31 g,86%). MH⁺=303. ¹H NMR (DMSO-d₆) 9.5 (s, 1H), 7.3 (m, 4H), 7.1 (t, 1H),6.35 (t, 1H), 5.35 (2d, 2H), 4.85 (d, 1H), 4.55 (d, 1H), 3.45 (m, 2H),3.2 (t, 1H), 3.0 (t, 1H), 2.05 (br d, 1H).

Compounds of the present invention can also be made by one skilled inthe art using the following general schemes:

Salts, Solvates and Prodrugs

Compounds of the present invention include pharmaceutically acceptablesalts, solvates and pro-drugs of the compounds disclosed herein.Pharmaceutically acceptable salts include salts derived from inorganicbases such as Li, Na, K, Ca, Mg, Fe, Cu, Zn, Mn; salts of organic basessuch as N,N′-diacetylethylenediamine, glucamine, triethylamine, choline,hydroxide, dicyclohexylamine, metformin, benzylamine, trialkylamine,thiamine; chiral bases like alkylphenylamine, glycinol, phenyl glycinol,salts of natural amino acids such as glycine, alanine, valine, leucine,isoleucine, norleucine, tyrosine, cystine, cysteine, methionine,proline, hydroxy proline, histidine, ornithine, lysine, arginine,serine; non-natural amino acids such as D-isomers or substituted aminoacids; guanidine, substituted guanidine wherein the substituents areselected from nitro, amino, alkyl, alkenyl, alkynyl, ammonium orsubstituted ammonium salts and aluminum salts. Salts may include acidaddition salts where appropriate which are, hydrochlorides, sulphates,nitrates, phosphates, perchlorates, borates, hydrohalides, acetates,tartrates, maleates, citrates, succinates, palmoates,methanesulphonates, benzoates, salicylates, benzenesulfonates,ascorbates, glycerophosphates, ketoglutarates. In one embodiment, thepharmaceutically acceptable salt of the compounds disclosed herein isthe hydrochloride salt.

“Solvate” denotes a physical association of a compound with one or moresolvent molecules. This physical association involves varying degrees ofionic and covalent bonding, including hydrogen bonding. In certaininstances the solvate will be capable of isolation, for example when oneor more solvent molecules are incorporated in the crystal lattice of thecrystalline solid. “Solvate” encompasses both solution-phase andisolatable solvates. “Hydrate” is a solvate wherein the solvent moleculeis H₂O. Other non-limiting examples of suitable solvates includealcohols (e.g., ethanolates, methanolates, and the like).

Prodrugs are compounds which are converted in vivo to active forms (see,e.g., R. B. Silverman, 1992, “The Organic Chemistry of Drug Design andDrug Action”, Academic Press, Chapter 8, incorporated herein byreference). Additionally, a discussion of prodrugs is provided in T.Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Volume 14 ofthe A.C.S. Symposium Series, and in Bioreversible Carriers in DrugDesign, Edward B. Roche, ed., American Pharmaceutical Association andPergamon Press, 1987, both of which are incorporated herein by referencethereto. Prodrugs can be used to alter the biodistribution (e.g., toallow compounds which would not typically enter the reactive site of theprotease) or the pharmacokinetics for a particular compound. Forexample, a carboxylic acid group, can be esterified, e.g., with a methylgroup or an ethyl group to yield an ester. When the ester isadministered to a subject, the ester is cleaved, enzymatically ornon-enzymatically, reductively, oxidatively, or hydrolytically, toreveal the anionic group. An anionic group can be esterified withmoieties (e.g., acyloxymethyl esters) which are cleaved to reveal anintermediate compound which subsequently decomposes to yield the activecompound.

Examples of prodrugs and their uses are well known in the art (See,e.g., Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm. Sci.66:1-19). The prodrugs can be prepared in situ during the finalisolation and purification of the compounds, or by separately reactingthe purified compound with a suitable derivatizing agent. For examplehydroxy groups can be converted into esters via treatment with acarboxilic acid in the presence of a catalyst. Examples of cleavablealcohol prodrug moieties include substituted and unsubstituted, branchedor unbranched lower alkyl ester moieties, (e.g., ethyl esters), loweralkenyl esters, di-lower alkyl-amino lower-alkyl esters (e.g.,dimethylaminoethyl ester), acylamino lower alkyl esters, acyloxy loweralkyl esters (e.g., pivaloyloxymethyl ester), aryl esters (phenylester), aryl-lower alkyl esters (e.g., benzyl ester), substituted (e.g.,with methyl, halo, or methoxy substituents) aryl and aryl-lower alkylesters, amides, lower-alkyl amides, di-lower alkyl amides, and hydroxyamides.

All stereoisomers (for example, geometric isomers, optical isomers andthe like) of the compounds disclosed herein (including those of thesalts, solvates and prodrugs of these compounds as well as the salts andsolvates of the prodrugs), such as those which may exist due toasymmetric carbons on various substituents, including enantiomeric forms(which may exist even in the absence of asymmetric carbons), rotamericforms, atropisomers, and diastereomeric forms, are contemplated withinthe scope of this invention. Individual stereoisomers of these compoundsmay, for example, be substantially free of other isomers, or may beadmixed, for example, as racemates or with all other, or other selected,stereoisomers. The chiral centers of the aforementioned compounds canhave the S or R configuration as defined by the IUPAC 1974Recommendations. The use of the terms “salt”, “solvate” “prodrug” andthe like, is intended to equally apply to the salt, solvate and prodrugof enantiomers, stereoisomers, rotamers, tautomers, racemates orprodrugs of the compounds of the present invention disclosed herein.

Formulations

The therapeutic agent(s) can be formulated to be suitable for any routeof administration, including e.g., orally in the form of tablets orcapsules or liquid, or in sterile aqueous solution for injection. Whenthe therapeutic agent(s) is formulated for oral administration, tabletsor capsules can be prepared by conventional means with pharmaceuticallyacceptable excipients such as binding agents (e.g., pregelatinized maizestarch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers(e.g., lactose, microcrystalline cellulose or calcium hydrogenphosphate); lubricants (e.g., magnesium stearate, talc or silica);disintegrants (e.g., potato starch or sodium starch glycolate); orwetting agents (e.g., sodium lauryl sulphate). The tablets may be coatedby methods well known in the art. Liquid preparations for oraladministration may take the form of, for example, solutions, syrups orsuspensions, or they may be presented as a dry product for constitutionwith water or another suitable vehicle before use. Such liquidpreparations may be prepared by conventional means with pharmaceuticallyacceptable additives such as suspending agents (e.g., sorbitol syrup,cellulose derivatives or hydrogenated edible fats); emulsifying agents(e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oilyesters, ethyl alcohol or fractionated vegetable oils); or preservatives(e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid). The liquidpreparations may also contain buffer salts, flavoring, coloring orsweetening agents as appropriate. Preparations for oral administrationmay be suitably formulated to give controlled or sustained release ofthe therapeutic agent(s).

In certain embodiments of the present invention, the therapeuticagent(s) is administered in a dosage form that permits systemic uptake,such that the therapeutic agent(s) may cross the blood-brain barrier soas to exert effects on neuronal cells. For example, pharmaceuticalformulations of the therapeutic agent(s) suitable forparenteral/injectable use generally include sterile aqueous solutions(where water soluble), or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersion. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol,polyethylene glycol, and the like), suitable mixtures thereof, orvegetable oils. The proper fluidity can be maintained, for example, bythe use of a coating such as lecithin, by the maintenance of therequired particle size in the case of dispersion and by the use ofsurfactants. Prevention of the action of microorganisms can be broughtabout by various antibacterial and antifungal agents, for example,parabens, chlorobutanol, phenol, benzyl alcohol, sorbic acid, and thelike. In many cases, it will be reasonable to include isotonic agents,for example, sugars or sodium chloride. Prolonged absorption of theinjectable compositions can be brought about by the use in thecompositions of agents delaying absorption, for example, aluminummonosterate or gelatin.

Sterile injectable solutions are prepared by incorporating thetherapeutic agent(s) in the required amount in the appropriate solventwith various of the other ingredients enumerated above, as required,followed by filter or terminal sterilization. Generally, dispersions areprepared by incorporating the various sterilized active ingredients intoa sterile vehicle which contains the basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum drying and the freeze-dryingtechnique which yield a powder of the active ingredient plus anyadditional desired ingredient from previously sterile-filtered solutionthereof.

The formulation can contain an excipient. Pharmaceutically acceptableexcipients which may be included in the formulation are buffers such ascitrate buffer, phosphate buffer, acetate buffer, and bicarbonatebuffer, amino acids, urea, alcohols, ascorbic acid, phospholipids;proteins, such as serum albumin, collagen, and gelatin; salts such asEDTA or EGTA, and sodium chloride; liposomes; polyvinylpyrollidone;sugars, such as dextran, mannitol, sorbitol, and glycerol; propyleneglycol and polyethylene glycol (e.g., PEG-4000, PEG-6000); glycerol;glycine or other amino acids; and lipids. Buffer systems for use withthe formulations include citrate; acetate; bicarbonate; and phosphatebuffers. Phosphate buffer is a preferred embodiment.

The formulation can also contain a non-ionic detergent. Preferrednon-ionic detergents include Polysorbate 20, Polysorbate 80, TritonX-100, Triton X-114, Nonidet P-40, Octyl a-glucoside, Octyl 8-glucoside,Brij 35, Pluronic, and Tween 20.

Routes of Administration

The therapeutic agent(s) may be administered orally or parenterally,including intravenously, subcutaneously, intra-arterially,intraperitoneally, ophthalmically, intramuscularly, buccally, rectally,vaginally, intraorbitally, intracerebrally, intradermally,intracranially, intraspinally, intraventricularly, intrathecally,intracisternally, intracapsularly, intrapulmonarily, intranasally,transmucosally, transdermally, or via inhalation. In one preferredembodiment, the therapeutic agent(s) is administered orally.

Administration of therapeutic agent(s) may be by periodic injections ofa bolus of the formulation, or may be administered by intravenous orintraperitoneal administration from a reservoir which is external (e.g.,an i.v. bag) or internal (e.g., a bioerodable implant). See, e.g., U.S.Pat. Nos. 4,407,957 and 5,798,113, each incorporated herein byreference. Intrapulmonary delivery methods and apparatus are described,for example, in U.S. Pat. Nos. 5,654,007, 5,780,014, and 5,814,607, eachincorporated herein by reference. Other useful parenteral deliverysystems include ethylene-vinyl acetate copolymer particles, osmoticpumps, implantable infusion systems, pump delivery, encapsulated celldelivery, liposomal delivery, needle-delivered injection, needle-lessinjection, nebulizer, aerosolizer, electroporation, and transdermalpatch. Needle-less injector devices are described in U.S. Pat. Nos.5,879,327; 5,520,639; 5,846,233 and 5,704,911, the specifications ofwhich are herein incorporated by reference. Any of the formulationsdescribed above can be administered using these methods.

Subcutaneous injections have the advantages allowingself-administration, while also resulting in a prolonged plasmahalf-life as compared to intravenous administration. Furthermore, avariety of devices designed for patient convenience, such as refillableinjection pens and needle-less injection devices, may be used with theformulations of the present invention as discussed herein.

Dosage

A suitable pharmaceutical preparation is in a unit dosage form. In suchform, the preparation is subdivided into suitably sized unit dosescontaining appropriate quantities of the active component, e.g., aneffective amount to achieve the desired purpose. In certain embodiments,the therapeutic agent(s) is administered in one or more daily doses(e.g., once-a-day, twice-a-day, thrice-a-day). In certain embodiments,the therapeutic agent(s) is administered in intermittently.

Exemplary dosing regimens are described in International patentapplication PCT/US08/61764 published as WO 2008/134628 on Jun. 11, 2008and U.S. provisional patent application 61/108,192, filed on Oct. 24,2008, both of which are incorporated by reference herein in theirentirety. In one embodiment, the therapeutic agent(s) is administered inan intermittent dosing regimen that includes an initial “loading dose”given daily, followed by a period of non-daily interval dosing.

The amount of effective therapeutic agent(s) for preventing or treatingthe referenced disorder can be determined on a case-by-case basis bythose skilled in the art. The amount and frequency of administration ofthe therapeutic agent(s) will be regulated according to the judgment ofthe attending clinician (physician) considering such factors as age,condition and size of the patient as well as risk for developingdisorder or severity of the symptoms of the referenced disorder beingtreated.

Combination Drug Therapy

The therapeutic agent(s) of the present invention can be administered incombination with at least one other therapeutic agent. Administration ofthe therapeutic agent(s) of the present invention with at least oneother therapeutic agent is understood to encompass administration thatis sequential or concurrent. In one embodiment, the therapeutic agentsare administered in separate dosage forms. In another embodiment, two ormore therapeutic agents are administered concurrently in the same dosageform.

In certain embodiments, the therapeutic agent(s) of the presentinvention are administered in combination with at least one othertherapeutic agent which is an anti-dyskinesia Agent (e.g., Carbidopa,Levodopa), an anti-infective agent (e.g., Miglustat), an antineoplasticagent (e.g., Busulfan, Cyclophosphamide), a gastrointestinal agent(e.g., Methylprednisolone), a micronutrient (e.g., Calcitriol,Cholecalciferol, Ergocalciferols, Vitamin D), a vasoconstrictor agent(e.g., Calcitriol).

In certain embodiments, the therapeutic agent(s) of the presentinvention are administered in combination with allopregnanolone, alow-cholesterol diet, or cholesterol-lowering agents such as statins(e.g., Lipitor®); fibrates such as fenofibrate (Lipidil®); niacin;and/or binding resins such as cholestyramine (Questran®).

In one embodiment, the therapeutic agent(s) of the present invention isadministered in combination with gene therapy. Gene therapy iscontemplated both with replacement genes such as glucocerebrosidase orwith inhibitory RNA (siRNA) for the SNCA gene. Gene therapy is describedin more detail in U.S. Pat. No. 7,446,098, filed on Feb. 17, 2004.

In one embodiment, the therapeutic agent(s) of the present invention isadministered in combination with at least one other therapeutic agentwhich is an anti-inflammatory agent (e.g., ibuprofen or other NSAID).

In one embodiment, the therapeutic agent(s) of the present invention isadministered in combination with a substrate inhibitor forglucocerebrosidase, such as N-butyl-deoxynojirimycin (Zavesca®;miglustat available from Actelion Pharmaceuticals, US, Inc., South SanFrancisco, Calif., US).

Combinations of the therapeutic agent(s) of the present invention withat least one other therapeutic agent which is a therapeutic agent forone or more other lysosomal enzymes are also contemplated. Following isa non-limiting list of therapeutic agents for lysosomal enzymes.

TABLE 1 LYSOSOMAL ENZYME THERAPEUTIC AGENT α-Glucosidase1-deoxynojirimycin (DNJ) GenBank Accession No. Y00839 α-homonojirimycincastanospermine Acid β-Glucosidase (β- isofagomine glucocerebrosidase)C-benzyl isofagomine and derivatives GenBank Accession No. J03059N-alkyl (C9-12)-DNJ Glucoimidazole (and derivatives) C-alkyl-IFG (andderivatives) N-alkyl-β-valeinamines Fluphenozine calystegines A₃, B₁, B₂and C₁ α-Galactosidase A 1-deoxygalactonojirimycin (DGJ) GenBankAccession No. NM000169 α-allo-homonojirimycin α-galacto-homonojirimycinβ-1-C-butyl-deoxynojirimycin calystegines A₂ and B₂ N-methylcalystegines A₂ and B₂ Acid β-Galactosidase 4-epi-isofagomine GenBankAccession No. M34423 1-deoxygalactonojirimyicn Galactocerebrosidase(Acid β- 4-epi-isofagomine Galactosidase) 1-deoxygalactonojirimycinGenBank Accession No. D25283 Acid α-Mannosidase 1-deoxymannojirimycinGenBank Accession No. U68567 Swainsonine Mannostatin A Acidβ-Mannosidase 2-hydroxy-isofagomine GenBank Accession No. U60337 Acidα-L-fucosidase 1-deoxyfuconojirimycin GenBank Accession No. NM_000147β-homofuconojirimycin 2,5-imino-1,2,5-trideoxy-L-glucitol2,5-deoxy-2,5-imino-D-fucitol 2,5-imino-1,2,5-trideoxy-D-altritolα-N-Acetylglucosaminidase 1,2-dideoxy-2-N-acetamido-nojirimycin GenBankAccession No. U40846 α-N-Acetylgalactosaminidase1,2-dideoxy-2-N-acetamido- GenBank Accession No. M62783galactonojirimycin β-Hexosaminidase A 2-N-acetylamino-isofagomineGenBank Accession No. NM_000520 1,2-dideoxy-2-acetamido-nojirimycinNagstatin β-Hexosaminidase B 2-N-acetamido-isofagomine GenBank AccessionNo. NM_000521 1,2-dideoxy-2-acetamido-nojirimycin Nagstatinα-L-Iduronidase 1-deoxyiduronojirimycin GenBank Accession No. NM_0002032-carboxy-3,4,5-trideoxypiperidine β-Glucuronidase 6-carboxy-isofagomineGenBank Accession No. NM_000181 2-carboxy-3,4,5-trideoxypiperidineSialidase 2,6-dideoxy-2,6, imino-sialic acid GenBank Accession No.U84246 Siastatin B Iduronate sulfatase 2,5-anhydromannitol-6-sulphateGenBank Accession No. AF_011889 Acid sphingomyelinase desipramine,phosphatidylinositol-4,5- GenBank Accession No. M59916 diphosphate

In certain embodiments, the therapeutic agent(s) of the presentinvention are administered in combination with at least one therapeuticagent which is an anti-dyskinesia Agent (e.g., Carbidopa, Levodopa), ananti-infective agent (e.g., Cyclosporine, Miglustat, Pyrimethamine), anantineoplastic agent (e.g., Alemtuzumab, Azathioprine, Busulfan,Clofarabine, Cyclophosphamide, Melphalan, Methotrexate, Rituximab), anantirheumatic agent (e.g., Rituximab) a gastrointestinal agent (e.g.,Methylprednisolone), a micronutrient (e.g., Calcitriol, Cholecalciferol,Ergocalciferols, Folic Acid, Vitamin D), a reproductive control agent(e.g., Methotrexate), a respiratory system agent (e.g.,Tetrahydrozoline), vasoconstrictor agent (e.g., Calcitriol,Tetrahydrozoline).

In certain embodiments, the therapeutic agent(s) of the presentinvention are administered in combination with at least one therapeuticagent which is a therapeutic agent for β-hexosaminidase A and/or atherapeutic agent for acid β-galactosidase. In certain embodiments, thetherapeutic agent(s) of the present invention are administered incombination with at least one therapeutic agent which is ananti-infective agent (e.g., Miglustat), an antineoplastic agent (e.g.,Alemtuzumab, Busulfan, Cyclophosphamide), a gastrointestinal agent(e.g., Methylprednisolone).

The therapeutic agent(s) of the present invention can be administered incombination with at least one other therapeutic agent which includes butis not limited to, RNAi, dopamine replacement (e.g., levadopa (L-DOPA)),dopamine replacement stabilizer (e.g., carbidopa, and entacapone),anticholinergic (e.g., trihexyphenidyl, benzotropine mesylate(Cogentin®), trihexyphenidyl HCL (Artane®), and procyclidine),catechol-O-methyltransferase (COMT) inhibitor (e.g., entacapone(Comtan®) and tolcapone (Tasmar®)), dopamine receptor agonist (e.g.,bromocriptine (Parlodel®), pramipexole (Mirapex®), ropinirole(Requip®)), pergolide (Permax), and APOKYN™ injection (apomorphinehydrochloride), monoamine oxidase (MAO) inhibitor (i.e., MAO-A and/orMAO-B inhibitors, e.g., selegiline (Deprenyl, Eldepryl®, Carbex®),selegiline HCl orally disintegrating tablet (Zelapar®), and rasagiline(Azilect®)), peripheral decarboxylase inhibitor, amantadine(Symmetrel®), and rivastigmine tartrate (Exelon®).

Also contemplated are combinations of the therapeutic agent(s) of thepresent invention with more than one other therapeutic agent. Exemplarycombinations of other therapeutic agents include, but not are notlimited to, carbidopa/levodopa (Sinemet® or Parcopa®), carbidopa,levodopa and entacapone (Stalevo®), levodopa with a dopamine receptoragonist such as bromocriptine (Parlodel®), pramipexole (Mirapex®),ropinirole (Requip®)), pergolide (Permax), or APOKYN™ injection(apomorphine hydrochloride).

In one embodiment, the therapeutic agent(s) of the present invention isadministered in combination with vaccine therapy, such as a vaccinecomprising alpha-synuclein and an adjuvant (Pilcher et al., LancetNeurol. 2005; 4(8):458-9).

In one embodiment, the therapeutic agent(s) of the present invention isadministered in combination with at least one other therapeutic agentthat may be protective such as dextromethorphan (Li et al., FASEB J.2005; April; 19(6):489-96); genistein (Wang et al., Neuroreport. 2005;February 28; 16(3):267-70), or minoclycline (Blum et al., Neurobiol Dis.2004; December; 17(3):359-66).

In one embodiment, the therapeutic agent(s) of the present invention isadministered in combination with at least one other therapeutic agentwhich is therapeutic agent for alpha-synuclein (e.g., Hsp70).

Patients having Parkinson's disease experience tremor, rigidity,bradykinesia, and postural imbalance. Patients having Lewy Body Dementiaexperience strong psychotic symptoms (visual hallucinations) in additionto mental decline such as memory loss and an inability to carry outsimple tasks. Observable improvements in symptoms, or a delay of onsetof certain symptoms in patients at risk of developing a disorder, or adelay in progression of the disorder will be evidence of a favorableresponse to the therapies provided herein.

In addition, measurable surrogate markers also may be useful forevaluating response to therapy. For instance, some investigators havereported detecting higher levels of alpha-synuclein or oligomeric formsof alpha-synuclein have been detected in the plasma of patients withParkinson's disease (Lee et al., J Neural Transm. 2006; 113(10):1435-9;El-Agnaf et al., FASEB J. 2006; 20(3):419-25), while some have reporteddecreased plasma alpha-synuclein in Parkinson's patients compared withnormal controls (Li et al., Exp Neurol. 2007; 204(2):583-8).

In certain embodiments, the therapeutic agent(s) of the presentinvention is administered in combination with at least one othertherapeutic agent which is an alcohol deterrent (e.g., Acamprosate), anarcotic analgesic (e.g., Remifentanil), an anti-dyskinesia agent (e.g,Amantadine, Apomorphine, Benserazide, Bromocriptine, Cabergoline,Carbidopa, Dexetimide, Droxidopa, Entacapone, Levodopa, Lisuride,Memantine, Piribedil, Pramipexol, Ropinirole, Selegiline, Sinemet), ananti-infective agent (e.g, Amantadine, Amoxicillin, Clarithromycin,Ethanol, Interferons, Minocycline, PS-K), an anti-obesity agent (e.g.,Phenylpropanolamine, Topiramate), an anticonvulsant (e.g., Etiracetam,Topiramate), an antiemetic (e.g., Trimethobenzamide), anantihypertensive agent (e.g., Trandolapril), an antineoplastic agent(e.g., Cabergoline, PS-K), central nervous system depressant (e.g.,Aripiprazole, Benzocaine, Clozapine, Cocaine, Dexmedetomidine,Diphenhydramine, Isoflurane, Lithium, Lithium Carbonate, Methylperon,Morphine, Propofol, Quetiapine, Raclopride, Remifentanil, SodiumOxybate), a central nervous system stimulant (e.g., Caffeine citrate,Modafinil, Nicotine polacrilex), a coagulant (e.g., ArginineVasopressin, Deamino Arginine Vasopressin, Vasopressins), a dermatologicagent (e.g., Loratadine, Promethazine), a gastrointestinal agent (e.g.,Diphenhydramine, Domperidone, Omeprazole, Trimethobenzamide), a hypnoticand/or sedative (e.g., Remifentanil), a micronutrient (e.g.,Alpha-Tocopherol, Coenzyme Q10, Ergocalciferols, Hydroxocobalamin, Iron,Tocopherol acetate, Tocopherols, Vitamin B 12, Vitamin D, Vitamin E), aneuroprotective agent (e.g., Eliprodil, Modafinil, Rasagiline,Rivastigmine, Topiramate), a nootropic agent (e.g., Donepezil,Etiracetam), a psychotropic drug (e.g., Aripiprazole, Citalopram,Clozapine, Duloxetine, Lithium, Lithium Carbonate, Methylperon,Nortriptyline, Paroxetine, Quetiapine, Raclopride, Venlafaxine), arespiratory system agent (e.g., Dextromethorphan, Guaifenesin,Ipratropium, Naphazoline, Oxymetazoline, Phenylephrine,Phenylpropanolamine), a vasoconstrictor agent (e.g., Naphazoline,Oxymetazoline, Phenylephrine, Phenylpropanolamine).

In one preferred embodiment, the aforementioned other therapeutic agentsare administered when the disorder is Parkinson's disease.

In certain embodiments, the therapeutic agent(s) of the presentinvention is administered in combination with at least one othertherapeutic agent which is a nicotinic alpha-7 agonist (e.g., MEM 3454or MEM 63908 both of which are available from Memory Pharmaceuticals).In certain embodiments, the therapeutic agent(s) of the presentinvention is administered in combination with at least one othertherapeutic agent which is R3487 and/or R4996 (both of which areavailable from Roche). Also contemplated are combinations of thetherapeutic agent(s) of the present invention with more than one othertherapeutic agents. Exemplary combinations of other therapeutic agentsinclude, but not are not limited to, R3487/MEM 3454 and R4996/MEM 63908.

In certain embodiments, the therapeutic agent(s) of the presentinvention is administered in combination with at least onecholinesterase inhibitor (e.g., donepezil (brand name Aricept),galantamine (brand name Razadyne), and rivastigmine (branded as Exelonand Exelon Patch).

In certain embodiments, the therapeutic agent(s) of the presentinvention is administered in combination with at least onenoncompetitive NMDA receptor antagonist (e.g., memantine (brand namesAkatinol, Axura, Ebixa/Abixa, Memox and Namenda)).

In certain embodiments, the therapeutic agent(s) of the presentinvention is administered in combination with at least one othertherapeutic agent which is a non-narcotic analgesic (e.g., Celecoxib,Resveratrol, Rofecoxib, TNFR-Fc fusion protein), an anti-dyskinesiaagent (e.g., Dexetimide, Gabapentin, Levodopa, Memantine), ananti-infective agent (e.g., Acetylcysteine, Acyclovir, Benzoates,Deoxyglucose, Doxycycline, Interferon Alfa-2a, Interferon-alpha,Interferons, Moxifloxacin, PS-K, Quinacrine, Rifampin, Salicylic Acid,Valacyclovir), an anti-Inflammatory agent (e.g., Aspirin, Celecoxib,Curcumin, Ibuprofen, Indomethacin, Naproxen, Resveratrol, Rofecoxib,TNFR-Fc fusion protein), an anti-obesity agent (e.g.,Phenylpropanolamine), an anticonvulsant agent (e.g., Gabapentin,Homotaurine, Lamotrigine), an antiemetic (e.g., Olanzapine), anantihypertensive agent (e.g., Trandolapril), an antilipemic agent (e.g.,Atorvastatin, Choline, Clofibric Acid, Pravastatin, Simvastatin), anantineoplastic agent (e.g., Bryostatin 1, Carmustine, Cyclophosphamide,Interferon Alfa-2a, Leuprolide, Medroxyprogesterone 17-Acetate,Methyltestosterone, PK 11195, Prednisone, PS-K, Resveratrol,2,3-dihydro-1H-imidazo[1,2-b]pyrazole), an antirheumatic agent (e.g.,Aspirin, Celecoxib, Curcumin, Ibuprofen, Indomethacin, Naproxen,Resveratrol, Rofecoxib, TNFR-Fc fusion protein), a central nervoussystem depressant (e.g., Aripiprazole, Benzocaine, Cocaine, Gabapentin,Haloperidol, Haloperidol decanoate, Lithium, Lithium Carbonate,Lorazepam, Midazolam, Olanzapine, Perphenazine, Propofol, Quetiapine,Risperidone, Sodium Oxybate, Trazodone, Valproic Acid, Zolpidem), acentral nervous system stimulant (e.g., Caffeine citrate, Modafinil,Nicotine polacrilex), a channel blocker (e.g., Gabapentin, Lamotrigine),a coagulant (e.g., Antiplasmin, Vitamin K), a dermatologic agent (e.g.,Mineral Oil, Salicylic Acid), a gastrointestinal agent (e.g., Choline,Haloperidol, Lorazepam, Olanzapine, Omeprazole, TNFR-Fc fusion protein),a hypnotic and/or sedative agent (e.g., Zolpidem), a hypoglycemic agent(e.g., Insulin, Asp(B28)-, Rosiglitazone), a micronutrient (e.g.,Alpha-Tocopherol, Ascorbic Acid, Coenzyme Q10, Copper, Folic Acid,Hydroxocobalamin, Inositol, Iron, Niacin, Niacinamide, Nicotinic Acids,Pyridoxine, Selenium, Thioctic Acid, Tocopherol acetate, Tocopherols,Vitamin B 12, Vitamin B 6, Vitamin E, Vitamin K), a neuroprotectiveagent (e.g., Huperzine A, Modafinil, Nefiracetam, Rasagiline,Rivastigmine, (3-aminopropyl)(n-butyl)phosphinic acid), a nootropicagent (e.g., Donepezil, Nefiracetam), a platelet aggregation inhibitor(e.g., Resveratrol), a psychotropic drug (e.g., Aripiprazole, Bupropion,Citalopram, Duloxetine, Gabapentin, Haloperidol, Haloperidol decanoate,Lithium, Lithium Carbonate, Lorazepam, Midazolam, Nefiracetam,Olanzapine, Paroxetine, Perphenazine, Quetiapine, Risperidone,Sertraline, Trazodone, Tryptophan, Valproic Acid, Venlafaxine), areproductive control agent (e.g., Estradiol 17 beta-cypionate, Estradiol3-benzoate, Estradiol valerate, Indomethacin, Leuprolide,Medroxyprogesterone, Medroxyprogesterone 17-Acetate, Mifepristone), arespiratory system agent (e.g., Acetylcysteine, Dextromethorphan,Guaifenesin, Naphazoline, Oxymetazoline, Phenylephrine,Phenylpropanolamine), or a vasoconstrictor agent (e.g., Naphazoline,Oxymetazoline, Phenylephrine, Phenylpropanolamine).

In one preferred embodiment, the aforementioned other therapeutic agentsare administered when the disorder is Alzheimer's disease.

EXAMPLES

The present invention is further described by means of the examples,presented below. The use of such examples is illustrative only and in noway limits the scope and meaning of the invention or of any exemplifiedterm. Likewise, the invention is not limited to any particular preferredembodiments described herein. Indeed, many modifications and variationsof the invention will be apparent to those skilled in the art uponreading this specification. The invention is therefore to be limitedonly by the terms of the appended claims along with the full scope ofequivalents to which the claims are entitled.

Example 1 Determination of Inhibition Constants

The binding affinity (defined here by K_(i) binding constant) of GCasefor novel compounds of the present invention were empirically determinedusing enzyme inhibition assays. In brief, the enzyme inhibition assaysused monitored the ability of a test compound to bind and prevent thehydrolysis of a fluorogenic substrate in a concentration-dependentmariner. Specifically, the enzyme activity of recombinant human GCase(rhGCase; Cerezymee, Genzyme Corp.) was measured using the4-methylumbelliferyl-β-D-glucopyranoside (4-MU-β-D-Glc) fluorogenicsubstrate in the absence or in the presence of varying amounts of eachtest compound. The resultant data were analyzed by comparing all testsamples to the no inhibition control sample (no compound; correspondingto 100% enzyme activity) to determine the residual enzyme activity inthe presence of test compound. The normalized residual activity datawere subsequently graphed (on y-axis) relative to the concentration oftest compound (on x-axis) to extrapolate the test compound concentrationwhich leads to 50% inhibition of enzyme activity (defined as IC₅₀). TheIC₅₀ value for each test compound was then inserted into theCheng-Prusoff equation (detailed below) to derive the absoluteinhibition constant K_(i) that accurately reflects the binding affinityof GCase for the test compound. The enzyme inhibition assays wereperformed at both pH 7.0 (endoplasmic reticulum pH) and at pH 5.2(lysosomal pH) to gain insight into the binding affinity (i.e., potency)of compounds for GCase in the endoplasmic reticulum and lysososome.

In Vitro Assay

Various concentrations of test compounds were prepared in buffer “M”consisting of 50 mM sodium phosphate buffer with 0.25% sodiumtaurocholate at pH 7.0 and pH 5.2. Enzyme (Cerezyme®, a recombinant formof the human enzyme β-glucocerebrosidase) was also diluted in the samebuffer “M” at pH 7.0 and pH 5.2. The substrate solution consisted of 3mM 4-methylumbelliferone β-D-glucopyranoside in buffer “M” with 0.15%Triton X-100 at both pH's. Five microliters of diluted enzyme was addedto 15 μl of the various inhibitor concentrations or buffer “M” alone andincubated at 37° C. for 1 hour with 50 μl of the substrate preparationto assess β-glucosidase activity at pH 7.0 and pH 5.2. Reactions werestopped by addition of an equal volume of 0.4 M glycine, pH 10.6.Fluorescence was measured on a plate reader for 1 sec/well using 355 nmexcitation and 460 nm emission. Incubations without added enzyme orwithout added inhibitors were used to define no enzyme activity andmaximum activity, respectively, and normalize % inhibition for a givenassay. The results of such in vitro inhibition assays for referencecompound IFG-tartrate and several test compounds are summarized below inTable 2A

TABLE 2A In vitro Determination of Inhibition Constants Cmpd CompoundIC₅₀ (μM) K_(i) (μM) IC₅₀ (μM) K_(i) (μM) # Name pH 5.2 pH 5.2 pH 7.0 pH7.0 6 (3R,4R,5S)-5- 0.0259 ± 0.0014 0.0136 ± 0.0008  0.0058 ± 0.000230.00306 ± 0.00012  (difluoromethyl)- piperidine-3,4-diol 13(3R,4R,5S)-5-(1- 0.0946 ± 0.0028 0.0498 ± 0.0015  0.0171 ± 0.0008  0.009± 0.0004 fluoroethyl)-piperidine- 3,4-diol* 9 (3R,4R,5R)-5-(1-  0.107 ±0.0041 0.044 ± 0.0017 0.020 ± 0.0008 0.010 ± 0.0004 hydroxyethyl)-piperidine-3,4-diol* 10 (3R,4R,5R)-5-(1- 0.343 ± 0.021 0.142 ± 0.00880.066 ± 0.0041 0.035 ± 0.0021 hydroxyethyl)- piperidine-3,4-diol* 14(3R,4R,5S)-5-(1-  0.038 ± 0.0016 0.016 ± 0.0007 0.007 ± 0.0003 0.004 ±0.0001 fluoroethyl)-piperidine- 3,4-diol* none (3R,4R,5S)-5-((R)-1-0.291 ± 0.006 0.121 ± 0.0026 0.060 ± 0.0029 0.031 ± 0.0015fluoropropyl)-piperidine- 3,4-diol hydrochloride none(3R,4R,5S)-5-benzyl- 0.659 ± 0.028 0.273 ± 0.012  0.127 ± 0.01  0.067 ±0.005  piperidine-3,4-diol none (3R,4R,5R)-5-((S)- 3.29 ± 0.25 1.36 ±0.10  0.017 ± 0.0035 0.0089 ± 0.0018  hydroxy(phenyl)methyl)-piperidine-3,4-diol none (3R,4R,5S)-5-(2-  0.234 ± 0.0037 0.097 ± 0.00150.029 ± 0.0013 0.015 ± 0.0007 hydroxypropan-2- yl)piperidine-3,4-diolnone IFG-tartrate  0.049 ± 0.0029 0.026 ± 0.0015 0.0074 ± 0.00007 0.0039 ± 0.000037 Notes: *Stereoisomer A and/or B

In Situ Assay

The effect of the novel compounds of the present invention on lysosomalGCase activity was assayed in situ using fibroblasts established from anormal subject. Cells seeded in 48-well plates were incubated with theindicated concentrations of compound for 16-24 hours. For thedose-response assays, cells were incubated with the in situ substrate5-(pentafluorobenzoylamino)fluorescein di-β-D-glucopyranoside(PFBFDβGlu) for 1 hour and subsequently lysed to determine the extent ofsubstrate hydrolysis in the presence of compound. The assay employed arange of 12 concentrations encompassing 5 orders of magnitude, centeredon the IC50. Specifically, the following concentration ranges wereemployed: (3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5R)-5-(1-hydroxyethyl)-piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol: 1.0×10⁻³ to 3.0×10⁻⁹ M;(3R,4R,5R)-5-(1-hydroxyethyl)-piperidine-3,4-diol: 1.0×10⁻⁴ to 3.0×10⁻¹⁰M; and (3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol: 1.0×10⁻³ to3.0×10⁻¹¹ M; wherein compound was serially diluted 1:3 from the highestconcentration in the ranges specified. Inhibition was determined as theratio of activity in the presence of compound to that in the absence ofcompound. For the washout assays, cells were treated with compound for16-24 hours at a concentration equal to the IC90. Cells were washedextensively and incubated in drug-free medium to allow net compoundefflux from cells. Cells were then tested for lysosomal GCase activityat 2 hour intervals over a total period of 8 hours following compoundremoval. The increase in activity over time was fitted with a singleexponential function to determine the compound's washout time. Theresults of these in situ inhibition assays for reference compoundIFG-tartrate and several test compounds are summarized below in Table2B.

TABLE 2B In situ Determination of Inhibition Constants Cmpd Compound Insitu IC₅₀ in situ # Name (μM) washout (hr) EC₅₀ (μM) E_(max) (%) 6(3R,4R,5S)-5- 0.408 ± 0.046 2.1 ± 0.30 0.018 ± 0.008 105.6 ± 8.7 (difluoromethyl)- piperidine-3,4-diol 13 (3R,4R,5S)-5-(1- 0.650 ± 0.1722.7 ± 0.12 0.044 ± 0.005 92.8 ± 6.6 fluoroethyl)-piperidine- 3,4-diol* 9(3R,4R,5R)-5-(1- 0.518 ± 0.022 10.5 ± 1.75  0.49 ± 0.06 83.7 ± 2.9hydroxyethyl)- piperidine-3,4-diol* 10 (3R,4R,5R)-5-(1- 0.798 ± 0.043 12 ± 1.65 1.06 ± 0.12 99.3 ± 4.9 hydroxyethyl)- piperidine-3,4-diol* 14(3R,4R,5S)-5-(1- 0.061 ± 0.019 3.7 ± 0.63 0.026 ± 0.003 89.7 ± 3.5fluoroethyl)-piperidine- 3,4-diol* none (3R,4R,5S)-5-((R)-1- 0.972 ±0.201 ND 0.086 ± 0.002 84.0 ± 4.1 fluoropropyl)-piperidine- 3,4-diolhydrochloride none (3R,4R,5S)-5-benzyl- 1.299 ± 0.323 1.2 ± 0.13 0.18 ±0.01 98.0 ± 4.5 piperidine-3,4-diol none (3R,4R,5R)-5-((S)- ND ND 4.99 ±0.86 72.1 ± 3.5 hydroxy(phenyl)methyl)- piperidine-3,4-diol none(3R,4R,5S)-5-(2- ND ND 0.791 ± 0.162 109.3 ± 3.6  hydroxypropan-2-yl)piperidine-3,4-diol none IFG-tartrate 0.271 ± 0.012 8.2 ± 0.04 0.31 ±0.11 105.5 ± 12.8 Notes: *Stereoisomer A and/or B Cheng-Prusoffequation: Ki = IC₅₀/(1 + [S]/K_(m)) where [S] = substrate concentration;2.5 mM 4-MU-β-D-Glc was used K_(m) = Michaelis constant that definessubstrate affinity; 1.8 ± 0.6 mM for 4-MU-β-D-Glc (Liou et al., (2006) JBiol. Chem. 281 (7), 4242-53)

When compared to reference compound IFG-tartrate, the following isnotable: (i) test compounds(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5R)-5-(1-hydroxyethyl)-piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol, were found to cause aconcentration-dependent increase in GCase activity and enhanced enzymeactivity to the same maximum level as reference compound IFG-tartrate atmuch lower concentration; (ii) test compounds(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol, washed out of the lysosomalcompartment (in situ washout) considerably faster than referencecompound IFG-tartrate; and (iii), test compounds(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5R)-5-(1-hydroxyethyl)-piperidine-3,4-diol,(3R,4R,5R)-5-(1-hydroxyethyl)-piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol, inhibited GCase activity.

Example 2 Blood Brain Barrier Penetration

The blood-brain barrier (BBB) penetration of reference compoundIFG-tartrate and several compounds of the present invention (i.e.,(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5R)-5-(1-hydroxyethyl)-piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol) were assayed after oraladministration to mice. For this purpose, 8-week old wild-type male mice(C57BU6) were administered a single 30 mg/kg (free base equivalent) p.o.dose of reference or test compound by gavage (n=3 mice per time point).Dosing solutions were prepared in water. After dosing, mice wereeuthanized with CO2 at the following time points: 0-, 0.5-, 1-, and4-hour post-dose. After euthanization, whole blood was collected fromthe inferior vena cava into lithium heparin tubes. Similarly, brainswere collected from each mouse. Plasma was derived by spinning wholeblood at 2,700×g for 10 minutes at 4° C. followed by storage on dry ice.Whole brains were washed in cold PBS to remove contaminating blood,blotted dry, flash frozen on dry ice, and ultimately stored at −80° C.until analysis. To prepare brain samples for analysis, 50-100 mg oftissue was homogenized in 400 μl of water/mg tissue. Samples were thenclarified by centrifugation. Next, 25 μl of the brain homogenatesupernatant or 25 μl of plasma were combined with 25 μl ofacetonitrile:water (95/5). This was supplemented with 25 μl ofacetonitrile and 50 μL of internal standard (100 ng/mL IFG-tartrate13C2-15N in 0.5% formic acid in (70:30) acetonitrile:methanol). Sampleswere again clarified by centrifugation and 75 μl of the supernatant wascombined with 75 μl of acetonitrile. Samples were then analyzed forcompound levels by LC-MS/MS at PPD Inc. (3230 Deming Way, Middleton,Wis. 53562). In brief, a Thermo Betasil, Silica-100, 50×3 mm, 5μ columnequilibrated with a mixture of mobile phase consisting of 5 mM ammoniumformate and 0.05% formic acid in (A) 95:5 acetonitrile:water or (B)70:20:10 methanol:water:acetonitrile was employed. Between 20 and 30 μlsample was injected for analysis. For calculating drug concentrations,raw data for plasma (ng/mL) and brain (ng/g) was converted to nM usingthe molecular weight of respective compounds and assuming 1 g of tissueis equivalent to 1 mL volume. Concentration as a function of time wasplotted in GraphPad Prism version 4.02.

The plasma levels and brain levels detected in mice administered asingle 30 mg/kg (free base equivalent) p.o. dose of reference compound(I.e., IFG-tartrate) or test compound (i.e.,(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5R)-5-(1-hydroxyethyl)-piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, or(3R,4R,5S)-5-benzylpiperidine-3,4-diol) reflect that(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol crossed the blood brain barriermore readily as compared to IFG-tartrate. Additionally, higher levels of(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol were detected in brain than thatobserved following administration of IFG-tartrate.

Example 3 GCase Enhancement

The ability of orally administered test compounds((3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, or(3R,4R,5S)-5-benzylpiperidine-3,4-diol) to elevate GCase levels wasassessed in mice. For this purpose, 8-week old wild-type male mice(C57BL/6) were administered a single p.o. (gavage) dose of a compound ofthe present invention (i.e.,(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol,(3R,4R,5S)-5-((R)-1-fluoropropyl)piperidine-3,4-diol hydrochloride, or(3R,4R,5S)-5-benzylpiperidine-3,4-diol). Details of the doseadministered for each compound are provided in Tables 3A and 3B. Thedosing solutions were prepared in water. Compounds were administeredover 2 weeks as follows: week 1, Mon-Fri (On), Sat-Sun (Off); week 2,Mon-Thu (On); necropsy on Friday. Thus, a total of 9 doses (dosingsolutions prepared fresh every day) were given to each mouse, with a24-hour washout between the last dose and necropsy.

After completion of dosing, mice were euthanized with CO2 and wholeblood was drawn into lithium heparin tubes from the inferior vena cava.Plasma was collected by spinning blood at 2700 g for 10 minutes at 4° C.Liver, spleen, lung, and brain tissues were removed, washed in cold PBS,blotted dry, flash frozen on dry ice, and stored at −80° C. untilanalysis. GCase levels were measured by homogenizing approximately 50 mgtissue in 500 μL McIlvane (MI) buffer (100 mM sodium citrate, 200 mMsodium phosphate dibasic, 0.25% sodium taurocholate, and 0.1% TritonX-100, pH 5.2) at pH 5.2 for 3-5 seconds on ice with a microhomogenizer. Homogenates were then incubated at room temperature withoutand with 2.5 mM conduritol-B-epoxide (CBE) for 30 min. Finally, 3.7 mM4-methylumbeliferryl-β-glucoside (4-MUG) substrate was added andincubated at 37° C. for 60 min. Reactions were stopped by addition of0.4 M glycine, pH 10.6. Fluorescence was measured on a plate reader for1 sec/well using 355 nm excitation and 460 nm emission. Total proteinwas determined in lysates using the MicroBCA kit according to themanufacturer's instructions. A 4-methylumbelliferone (4-MU) standardcurve ranging from 1.0 nM to 50 μM was run in parallel for conversion ofraw fluorescence data to absolute GCase activity (in the presence andabsence of CBE) and expressed as nanomoles of 4-MU released permilligram of protein per hour (nmol/mg protein/hr). GCase levels andprotein levels were calculated using Microsoft Excel (Redmond, Wash.)and GraphPad Prism version 4.02.

Tables 3A and 3B summarize the dose administered for each compoundexamined in mice as described above as well as the resultant level ofGCase enhancement in brain and spleen, respectively, compoundconcentration in tissue, compound concentration in GCase assay andinhibition constant (Ki).

TABLE 3A GCase enhancement in Brain Compound Compound Dose GCaseconcentration concentration (mg/kg) increase in tissue2.2 in GCase Ki pHCompound Name FBE (fold) nmol/kg assay (μM) 5.2 (uM) (3R,4R,5S)-5- 102.1 55 0.0002 0.0136 ± 0.0008  (difluoromethyl)- piperidine-3,4-diol(3R,4R,5S)-5- 100 2.6 301  0.0010 (difluoromethyl)- piperidine-3,4-diol(3R,4R,5S)-5-(1- 10 1.5 50 0.0002 0.0498 ± 0.0015 fluoroethyl)-piperidine- 3,4-diol* (3R,4R,5S)-5-(1- 100 2.4 415  0.0014fluoroethyl)-piperidine- 3,4-diol* (3R,4R,5R)-5-(1- ND ND ND ND 0.044 ±0.0017 hydroxyethyl)- piperidine-3,4-diol* (3R,4R,5R)-5-(1- ND ND ND ND0.142 ± 0.0088 hydroxyethyl)- piperidine-3,4-diol* (3R,4R,5S)-5-(1- 101.5 BLQ(1) BLQ 0.016 ± 0.0007 fluoroethyl)-piperidine- 3,4-diol*(3R,4R,5S)-5-(1- 100 2.2 41 0.0001 fluoroethyl)-piperidine- 3,4-diol*(3R,4R,5S)-5-((R)-1- 10 0.9 BLQ(2) BLQ 0.121 ± 0.0026fluoropropyl)-piperidine- 3,4-diol hydrochloride (3R,4R,5S)-5-((R)-1-100 1.1 38 0.0001 fluoropropyl)-piperidine- 3,4-diol hydrochloride(3R,4R,5S)-5-benzyl- 10 1.2 ND ND 0.273 ± 0.012  piperidine-3,4-diol(3R,4R,5S)-5-benzyl- 100 1.4 ND ND piperidine-3,4-diol(3R,4R,5R)-5-((S)- ND ND ND ND 1.36 ± 0.10  hydroxy(phenyl)methyl)-piperidine-3,4-diol (3R,4R,5S)-5-(2- ND ND ND ND 0.097 ± 0.0015hydroxypropan-2- yl)piperidine-3,4-diol Notes: *Stereoisomer A and/or B(1)BLQ < 7.4 nmol/kg; (2)BLQ < 2.2 nmol/kg ND: Not determined

TABLE 3B GCase enhancement in Spleen Compound Dose GCase Compoundconcentration (mg/kg) increase concentration in GCase Ki pH CompoundName FBE (fold) in tissue assay 5.2 (uM) (3R,4R,5S)-5- 10 1.9 100 0.00030.0136 ± 0.0008  (difluoromethyl)- piperidine-3,4-diol (3R,4R,5S)-5- 1002.4 435 0.0015 (difluoromethyl)- piperidine-3,4-diol (3R,4R,5S)-5-(1- 101.0 BLQ(1) BLQ 0.0498 ± 0.0015  fluoroethyl)-piperidine- 3,4-diol*(3R,4R,5S)-5-(1- 100 1.5 948 0.0032 fluoroethyl)-piperidine- 3,4-diol*(3R,4R,5R)-5-(1- ND ND ND ND 0.044 ± 0.0017 hydroxyethyl)-piperidine-3,4-diol* (3R,4R,5R)-5-(1- ND ND ND ND 0.142 ± 0.0088hydroxyethyl)- piperidine-3,4-diol* (3R,4R,5S)-5-(1- 10 1.6 BLQ(2) BLQ0.016 ± 0.0007 fluoroethyl)-piperidine- 3,4-diol* (3R,4R,5S)-5-(1- 1002.3  99 0.0003 fluoroethyl)-piperidine- 3,4-diol* (3R,4R,5S)-5-((R)-1-10 0.7  21 0.0001 0.121 ± 0.0026 fluoropropyl)-piperidine- 3,4-diolhydrochloride (3R,4R,5S)-5-((R)-1- 100 0.7  60 0.0002fluoropropyl)-piperidine- 3,4-diol hydrochloride (3R,4R,5S)-5-benzyl- 101.0 ND ND 0.273 ± 0.012  piperidine-3,4-diol (3R,4R,5S)-5-benzyl- 1001.2 ND ND piperidine-3,4-diol (3R,4R,5R)-5-((S)- ND ND ND ND 1.36 ±0.10  hydroxy(phenyl)methyl)- piperidine-3,4-diol (3R,4R,5S)-5-(2- ND NDND ND 0.097 ± 0.0015 hydroxypropan-2- yl)piperidine-3,4-diol Notes:*Stereoisomer A and/or B (1)BLQ < 6.8 nmol/kg; (2)BLQ < 7.9 nmol/kg ND:Not determined

As reflected in Tables 3A and 3B, mice administered(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol, or(3R,4R,5S)-5-benzylpiperidine-3,4-diol demonstrated significant GCaseenhancement in brain and spleen.

Example 4 Rat Pharmacokinetics

Pharmacokinetic (PK) data was obtained in rats to assess thebioavailability of test compound. In particular, the following PKparameters were calculated: bioavailability as measured by area underthe Concentration/Time curve (AUC), fraction of dose available (% F;further defined below), clearance (CL), volume of distribution (Vd), andhalf-life (t1/2). For this purpose, 8-week old Sprague-Dawley male ratswere given either a single intravenous (IV) dose equivalent to 3 mg/kgof free base or single escalating p.o. (gavage) doses of test compoundequivalent to 10, 30, and 100 mg/kg of free base. Three rats were usedper dosing group. Blood was collected over a 24-hr period. The timepoints for blood collection after intravenous administration were: 0,2.5, 5, 10, 15, 30, 45 min, 1, 2, 4, 8, 12, and 24 hrs; time points forblood collection after p.o. administrations were: 0, 5, 15, 30, 45 min,1, 2, 3, 4, 8, 12, and 24 hrs. Plasma samples were analyzed for compoundlevels by LC-MS/MS at PPD. Raw data was analyzed by non-compartmentalanalysis in Win-nonLin to calculate V_(D), % F, CL, and t_(1/2).

Various pharmacokinetic parameters for(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol based on the aforementioned studyare detailed below in Tables 4A-D.

TABLE 4A Rat PK for (3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol-HClDose (mg/kg) Free AUC_(last) C_(max) Salt Base Route (hr * ng/ml) % Ft_(1/2) (h) (ng/mL) CL (mL/hr/kg) V_(D) (mL/kg)  3.65  3 IVAUC_(0-12 hr) N/A 1.1 ± 2323 ± 1555 ± 2612 ± 2044 ± 0.05 348 218 269 294 12.18  10 PO AUC_(0-12 hr) 106 ± 2.58 ± 3363 ± N/A N/A 6714 ± 8.6 0.78219 524  36.54  30 PO AUC_(0-24 hr) 101 ± 2.75 ± 10037 ± N/A N/A 21685 ±6.9 0.36 865 1515 121.81 100 PO AUC_(0-24 hr) 121± 2.41 ± 33200 ± N/AN/A 79389 ± 12.9 0.16 4990 8570 Notes: Non compartmental analysis meanvalues (N = 3 rats) BLD Below Limit of Detection (<0.5 ng/mL) BLQ BelowLimit of Quantitation${\% \mspace{14mu} F} = \frac{{AUC}\mspace{14mu} {PO} \times 100\mspace{14mu} {dose}\mspace{14mu} {normalized}}{{AUC}\mspace{14mu} {IV}}$AUC_(last) = Area under the Concentration/Time curve to the last datapoint

TABLE 4B Rat PK for (3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol-HCl*Dose (mg/kg) Free AUC_(last) C_(max) CL V_(D) Salt Base Route (hr*ng/ml)% F t_(1/2) (h) (ng/mL) (mL/hr/kg) (mL/kg) 3.67 3 IV AUC_(0-24 hr) N/A2.6 ± 0.64 2328 ± 373 2708 ± 410 9774 ± 1551   1421 ± 188.1 12.23 10 POAUC_(0-24 hr) 148 ± 12.5 2.8 ± 0.50 2680 ± 167 N/A N/A 7097 ± 606 36.7030 PO AUC_(0-24 hr) 155 ± 12.2 2.7 ± 0.12 6917 ± 451 N/A N/A 21664 ±1708 122.34 100 PO AUC_(0-24 hr) 142 ± 2.5  2.5 ± 0.19 19433 ± 3031 N/AN/A 59481 ± 1005 Note: *Stereoisomer A and/or B

TABLE 4C Rat PK for (3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol-HCl*Dose (mg/kg) Free AUC_(last) C_(max) CL V_(D) Salt Base Route (hr*ng/ml)% F t_(1/2) (h) (ng/mL) (mL/hr/kg) (mL/kg) 3.67 3 IV AUC_(0-24 hr) N/A2.06 ± 0.47   2427 ± 192 2188 ± 173 6304 ± 927  1370 ± 109 12.23 10 POAUC_(0-24 hr)  98 ± 1.85 3.0 ± 0.22 1127 ± 60 N/A N/A 4251 ± 88 36.70 30PO AUC_(0-24 hr) 104 ± 0.88 2.6 ± 0.16  4680 ± 369 N/A N/A 14229 ± 127122.34 100 PO AUC_(0-24 hr) 104 ± 1.5  2.4 ± 0.16 15733 ± 622 N/A N/A50946 ± 713 Note: *Stereoisomer A and/or B

TABLE 4D Rat PK for (3R,4R,5S)-5-benzylpiperidine-3,4-diol-HCl Dose(mg/kg) Free AUC_(last) C_(max) CL V_(D) Salt Base Route (hr*ng/ml) % Ft_(1/2) (h) (ng/mL) (mL/hr/kg) (mL/kg) 3.53 3 IV AUC_(0-12 hr) N/A 1.7 ±1.5  969 ± 104 5145 ± 532 12570 ± 1792  592 ± 60.9 11.76 10 POAUC_(0-24 hr) 61.7 ± 2.4 3.86 ± 0.6   641 ± 48.7 N/A N/A 1200 ± 46.435.28 30 PO AUC_(0-24 hr) 62.3 ± 1.2 3.8 ± 0.19 1703 ± 133  N/A N/A 3690± 71.5 117.59 100 PO AUC_(0-24 hr)  68.3 ± 10.8 2.9 ± 0.11 7140 ± 1357N/A N/A 13519 ± 2177 

As reflected in Tables 4A-D,(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol have favorable pharmacokineticprofiles for drug development. In particular,(3R,4R,5S)-5-(difluoromethyl)piperidine-3,4-diol,(3R,4R,5S)-5-(1-fluoroethyl)piperidine-3,4-diol, and(3R,4R,5S)-5-benzylpiperidine-3,4-diol show excellent oralbioavailability (approximately 50-100%) and dose proportionality, ahalf-life of 1.0 to 4.0 hours, and a volume of distribution suggestingadequate penetration into peripheral tissues.

1-22. (canceled)
 23. A compound of Formula II:

wherein: R¹ is C(R²)(R³)(R⁴); R² is hydrogen, —OH or halogen; R³ ishydrogen, —OH, halogen or —CH3; R⁴ is halogen, fluorophenyl,methylphenyl, cyclohexylmethyl; R³ and R⁴ may join with the carbon towhich they are attached to form a cycloalkyl ring, which may beoptionally substituted with one or more halogen atoms; R⁶ is hydrogen,phenylalkyl or substituted phenylalkyl; Z is optional, when present Z is—(CH₂)—, —C(═O) —S(═O)₂NH—, —S(═O)₂—, —S(═O)₂—CH₃, C(═O)—NH—,—S(═O)₂NR⁹R¹⁰, —C(═S)—NH— or —C(═O)—CH₃, R⁹ is hydrogen or CH₃; R¹⁰ ishydrogen or CH₃; R⁵ is hydrogen or aminophenylalkyl; R⁷ is —OH orhalogen; and R⁸ is hydrogen, halogen or —CH₃, provided that R² and R³cannot both be hydrogen when R⁴ is halogen, Z is not present, R⁷ is —OH,R⁵, R⁶ and R⁸ are hydrogen.
 24. A compound of Formula III:

wherein: R¹ is C(R²)(R³)(R⁴); R² is hydrogen, —OH or halogen; R³ ishydrogen, —OH, halogen or —CH₃; R⁴ is halogen, —CH₃, phenyl,fluorophenyl, methylphenyl, cyclohexylmethyl; R³ and R⁴ may join withthe carbon to which they are attached to form a cycloalkyl ring, whichmay be optionally substituted with one or more halogen atoms; R⁷ is —OHor halogen; and R⁸ is hydrogen, halogen or —CH₃, provided that R² and R³cannot both be hydrogen when R⁴ is a halogen, R⁷ is —OH and R⁸ ishydrogen.
 25. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt, or solvate thereof.
 26. Thecompound of claim 23 which is:

or a pharmaceutically acceptable salt thereof.
 27. The compound of claim23 which is:

or a pharmaceutically acceptable salt thereof.
 28. The compound of claim23 which is:

or a pharmaceutically acceptable salt thereof.
 29. The compound of claim23 which is:

or a pharmaceutically acceptable salt thereof.
 30. A pharmaceuticalcomposition comprising the compound of claim 1 and at least onepharmaceutically acceptable carrier.
 31. The pharmaceutical compositionof claim 30, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 32. The pharmaceuticalcomposition of claim 30, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 33. The pharmaceuticalcomposition of claim 30, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 34. The pharmaceuticalcomposition of claim 30, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 35. A pharmaceuticalcomposition comprising the compound of claim 2 and at least onepharmaceutically acceptable carrier.
 36. A pharmaceutical compositioncomprising the compound of claim 3 and at least one pharmaceuticallyacceptable carrier.